#METABOLOMICS WORKBENCH zhaom_20220406_222949 DATATRACK_ID:3178 STUDY_ID:ST002124 ANALYSIS_ID:AN003477 VERSION 1 CREATED_ON 05-31-2022 #PROJECT PR:PROJECT_TITLE Amino acid catabolism in hematopoietic cells PR:PROJECT_SUMMARY Hematopoietic stem cells (HSCs) adapt their metabolism to maintenance and PR:PROJECT_SUMMARY proliferation, but the mechanism remains incompletely understood. Here, we have PR:PROJECT_SUMMARY investigated the total levels, uptake and catabolism of amino acid in PR:PROJECT_SUMMARY hematopoietic stem cells, hemopoietic progenitors, and differentiated PR:PROJECT_SUMMARY hematopoietic cells. We have also studied the catabolism of amino acid in PR:PROJECT_SUMMARY hematopoietic stem cells under different conditions, such as homeostasis and PR:PROJECT_SUMMARY proliferation and with drug treatment. Moreover, glycolytic metabolite, NAD+ PR:PROJECT_SUMMARY precursor nicotinamide riboside (NR), accelerated AA catabolism to activate GCN2 PR:PROJECT_SUMMARY and sustain long-term function of HSCs. Overall, our study uncovers the direct PR:PROJECT_SUMMARY links between metabolic alterations and translation control in HSCs during PR:PROJECT_SUMMARY homeostasis and proliferation. PR:INSTITUTE Sun Yat-sen University PR:LAST_NAME Zhao PR:FIRST_NAME Meng PR:ADDRESS Zhongshan 2nd Road PR:EMAIL zhaom38@mail.sysu.edu.cn PR:PHONE 18138799889 PR:DOI http://dx.doi.org/10.21228/M8GD8D #STUDY ST:STUDY_TITLE Amino acids and TCA substrates in hematopoietic cells (Part1) ST:STUDY_SUMMARY This study uses [13C,15N] labeled amino acids to study the amino acid ST:STUDY_SUMMARY consumption and their catabolism into tricarboxylic acid cycle substrates in ST:STUDY_SUMMARY hematopoietic stem cells, hemopoietic progenitors, and differentiated ST:STUDY_SUMMARY hematopoietic cells under different conditions, such as homeostasis and ST:STUDY_SUMMARY proliferation and with drug treatment. ST:INSTITUTE Sun Yat-sen University ST:LAST_NAME Zhao ST:FIRST_NAME Meng ST:ADDRESS Zhongshan 2nd Road ST:EMAIL zhaom38@mail.sysu.edu.cn ST:PHONE 18138799889 ST:SUBMIT_DATE 2022-04-06 #SUBJECT SU:SUBJECT_TYPE Mammal SU:SUBJECT_SPECIES Mus musculus SU:TAXONOMY_ID 10090 #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data SUBJECT_SAMPLE_FACTORS - Lin–_1 Source_Name[gating]:Lineage-negative (Lin–) | Treatment:[13C, 15N] AAs were incorporated for 1 h | Batch:1a | Note:[13C, 15N] AAs were used for AA uptake (Fig1.B); unlabeled AAs were used for total AA (Fig1.A) and TCA substrate levels (Fig1.D). RAW_FILE_NAME=linn-1.cdf SUBJECT_SAMPLE_FACTORS - Lin–_2 Source_Name[gating]:Lineage-negative (Lin–) | Treatment:[13C, 15N] AAs were incorporated for 1 h | Batch:1a | Note:[13C, 15N] AAs were used for AA uptake (Fig1.B); unlabeled AAs were used for total AA (Fig1.A) and TCA substrate levels (Fig1.D). RAW_FILE_NAME=linn-2.cdf SUBJECT_SAMPLE_FACTORS - Lin–_3 Source_Name[gating]:Lineage-negative (Lin–) | Treatment:[13C, 15N] AAs were incorporated for 1 h | Batch:1a | Note:[13C, 15N] AAs were used for AA uptake (Fig1.B); unlabeled AAs were used for total AA (Fig1.A) and TCA substrate levels (Fig1.D). RAW_FILE_NAME=linn-3.cdf SUBJECT_SAMPLE_FACTORS - Lin–_0h_1 Source_Name[gating]:Lineage-negative (Lin–) | Treatment:[13C, 15N] AAs were incorporated for 24 h and subsequently washed out for 0 h | Batch:1b | Note:[13C, 15N] AA levels and [13C]TCA substrate levels in different time point were calculated for AA catabolism (Fig1.C). RAW_FILE_NAME=linn-0h-1.cdf SUBJECT_SAMPLE_FACTORS - Lin–_0h_2 Source_Name[gating]:Lineage-negative (Lin–) | Treatment:[13C, 15N] AAs were incorporated for 24 h and subsequently washed out for 0 h | Batch:1b | Note:[13C, 15N] AA levels and [13C]TCA substrate levels in different time point were calculated for AA catabolism (Fig1.C). RAW_FILE_NAME=linn-0h-2.cdf SUBJECT_SAMPLE_FACTORS - Lin–_0h_3 Source_Name[gating]:Lineage-negative (Lin–) | Treatment:[13C, 15N] AAs were incorporated for 24 h and subsequently washed out for 0 h | Batch:1b | Note:[13C, 15N] AA levels and [13C]TCA substrate levels in different time point were calculated for AA catabolism (Fig1.C). RAW_FILE_NAME=linn-0h-3.cdf SUBJECT_SAMPLE_FACTORS - Lin–_12h_1 Source_Name[gating]:Lineage-negative (Lin–) | Treatment:[13C, 15N] AAs were incorporated for 24 h and subsequently washed out for 12 h | Batch:1b | Note:[13C, 15N] AA levels and [13C]TCA substrate levels in different time point were calculated for AA catabolism (Fig1.C). RAW_FILE_NAME=linn-12h-1.cdf SUBJECT_SAMPLE_FACTORS - Lin–_12h_2 Source_Name[gating]:Lineage-negative (Lin–) | Treatment:[13C, 15N] AAs were incorporated for 24 h and subsequently washed out for 12 h | Batch:1b | Note:[13C, 15N] AA levels and [13C]TCA substrate levels in different time point were calculated for AA catabolism (Fig1.C). RAW_FILE_NAME=linn-12h-2.cdf SUBJECT_SAMPLE_FACTORS - Lin–_12h_3 Source_Name[gating]:Lineage-negative (Lin–) | Treatment:[13C, 15N] AAs were incorporated for 24 h and subsequently washed out for 12 h | Batch:1b | Note:[13C, 15N] AA levels and [13C]TCA substrate levels in different time point were calculated for AA catabolism (Fig1.C). RAW_FILE_NAME=linn-12h-3.cdf SUBJECT_SAMPLE_FACTORS - Lin–_6h_1 Source_Name[gating]:Lineage-negative (Lin–) | Treatment:[13C, 15N] AAs were incorporated for 24 h and subsequently washed out for 6 h | Batch:1b | Note:[13C, 15N] AA levels and [13C]TCA substrate levels in different time point were calculated for AA catabolism (Fig1.C). RAW_FILE_NAME=linn-6h-1.cdf SUBJECT_SAMPLE_FACTORS - Lin–_6h_2 Source_Name[gating]:Lineage-negative (Lin–) | Treatment:[13C, 15N] AAs were incorporated for 24 h and subsequently washed out for 6 h | Batch:1b | Note:[13C, 15N] AA levels and [13C]TCA substrate levels in different time point were calculated for AA catabolism (Fig1.C). RAW_FILE_NAME=linn-6h-2.cdf SUBJECT_SAMPLE_FACTORS - Lin–_6h_3 Source_Name[gating]:Lineage-negative (Lin–) | Treatment:[13C, 15N] AAs were incorporated for 24 h and subsequently washed out for 6 h | Batch:1b | Note:[13C, 15N] AA levels and [13C]TCA substrate levels in different time point were calculated for AA catabolism (Fig1.C). RAW_FILE_NAME=linn-6h-3.cdf SUBJECT_SAMPLE_FACTORS - HSC_1 Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:[13C, 15N] AAs were incorporated for 1 h | Batch:1a | Note:[13C, 15N] AAs were used for AA uptake (Fig1.B); unlabeled AAs were used for total AA (Fig1.A) and TCA substrate levels (Fig1.D). RAW_FILE_NAME=hsc-1.cdf SUBJECT_SAMPLE_FACTORS - HSC_2 Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:[13C, 15N] AAs were incorporated for 1 h | Batch:1a | Note:[13C, 15N] AAs were used for AA uptake (Fig1.B); unlabeled AAs were used for total AA (Fig1.A) and TCA substrate levels (Fig1.D). RAW_FILE_NAME=hsc-2.cdf SUBJECT_SAMPLE_FACTORS - HSC_3 Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:[13C, 15N] AAs were incorporated for 1 h | Batch:1a | Note:[13C, 15N] AAs were used for AA uptake (Fig1.B); unlabeled AAs were used for total AA (Fig1.A) and TCA substrate levels (Fig1.D). RAW_FILE_NAME=hsc-3.cdf SUBJECT_SAMPLE_FACTORS - HSC_Ctrl_0h_1 Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:HSC Basal, [13C, 15N] AAs were incorporated for 24 h and subsequently washed out for 0 h | Batch:1b | Note:[13C, 15N] AA levels and [13C]TCA substrate levels in different time point were calculated for AA catabolism (Fig1.C, Fig2.D). RAW_FILE_NAME=hsc-ct-0h-1.cdf SUBJECT_SAMPLE_FACTORS - HSC_Ctrl_0h_2 Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:HSC Basal, [13C, 15N] AAs were incorporated for 24 h and subsequently washed out for 0 h | Batch:1b | Note:[13C, 15N] AA levels and [13C]TCA substrate levels in different time point were calculated for AA catabolism (Fig1.C, Fig2.D). RAW_FILE_NAME=hsc-ct-0h-2.cdf SUBJECT_SAMPLE_FACTORS - HSC_Ctrl_0h_3 Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:HSC Basal, [13C, 15N] AAs were incorporated for 24 h and subsequently washed out for 0 h | Batch:1b | Note:[13C, 15N] AA levels and [13C]TCA substrate levels in different time point were calculated for AA catabolism (Fig1.C, Fig2.D). RAW_FILE_NAME=hsc-ct-0h-3.cdf SUBJECT_SAMPLE_FACTORS - HSC_Ctrl_12h_1 Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:HSC Basal, [13C, 15N] AAs were incorporated for 24 h and subsequently washed out for 12 h | Batch:1b | Note:[13C, 15N] AA levels and [13C]TCA substrate levels in different time point were calculated for AA catabolism (Fig1.C, Fig2.D). RAW_FILE_NAME=hsc-ct-12h-1.cdf SUBJECT_SAMPLE_FACTORS - HSC_Ctrl_12h_2 Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:HSC Basal, [13C, 15N] AAs were incorporated for 24 h and subsequently washed out for 12 h | Batch:1b | Note:[13C, 15N] AA levels and [13C]TCA substrate levels in different time point were calculated for AA catabolism (Fig1.C, Fig2.D). RAW_FILE_NAME=hsc-ct-12h-2.cdf SUBJECT_SAMPLE_FACTORS - HSC_Ctrl_12h_3 Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:HSC Basal, [13C, 15N] AAs were incorporated for 24 h and subsequently washed out for 12 h | Batch:1b | Note:[13C, 15N] AA levels and [13C]TCA substrate levels in different time point were calculated for AA catabolism (Fig1.C, Fig2.D). RAW_FILE_NAME=hsc-ct-12h-3.cdf SUBJECT_SAMPLE_FACTORS - HSC_Ctrl_6h_1 Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:HSC Basal, [13C, 15N] AAs were incorporated for 24 h and subsequently washed out for 6 h | Batch:1b | Note:[13C, 15N] AA levels and [13C]TCA substrate levels in different time point were calculated for AA catabolism (Fig1.C, Fig2.D). RAW_FILE_NAME=hsc-ct-6h-1.cdf SUBJECT_SAMPLE_FACTORS - HSC_Ctrl_6h_2 Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:HSC Basal, [13C, 15N] AAs were incorporated for 24 h and subsequently washed out for 6 h | Batch:1b | Note:[13C, 15N] AA levels and [13C]TCA substrate levels in different time point were calculated for AA catabolism (Fig1.C, Fig2.D). RAW_FILE_NAME=hsc-ct-6h-2.cdf SUBJECT_SAMPLE_FACTORS - HSC_Ctrl_6h_3 Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:HSC Basal, [13C, 15N] AAs were incorporated for 24 h and subsequently washed out for 6 h | Batch:1b | Note:[13C, 15N] AA levels and [13C]TCA substrate levels in different time point were calculated for AA catabolism (Fig1.C, Fig2.D). RAW_FILE_NAME=hsc-ct-6h-3.cdf SUBJECT_SAMPLE_FACTORS - LSK_1 Source_Name[gating]:Lin– Scal1+ c-Kit+ (LSK) | Treatment:[13C, 15N] AAs were incorporated for 1 h | Batch:1a | Note:[13C, 15N] AAs were used for AA uptake (Fig1.B); unlabeled AAs were used for total AA (Fig1.A) and TCA substrate levels (Fig1.D). RAW_FILE_NAME=lsk-1.cdf SUBJECT_SAMPLE_FACTORS - LSK_2 Source_Name[gating]:Lin– Scal1+ c-Kit+ (LSK) | Treatment:[13C, 15N] AAs were incorporated for 1 h | Batch:1a | Note:[13C, 15N] AAs were used for AA uptake (Fig1.B); unlabeled AAs were used for total AA (Fig1.A) and TCA substrate levels (Fig1.D). RAW_FILE_NAME=lsk-2.cdf SUBJECT_SAMPLE_FACTORS - LSK_3 Source_Name[gating]:Lin– Scal1+ c-Kit+ (LSK) | Treatment:[13C, 15N] AAs were incorporated for 1 h | Batch:1a | Note:[13C, 15N] AAs were used for AA uptake (Fig1.B); unlabeled AAs were used for total AA (Fig1.A) and TCA substrate levels (Fig1.D). RAW_FILE_NAME=lsk-3.cdf SUBJECT_SAMPLE_FACTORS - LSK_0h_1 Source_Name[gating]:Lin– Scal1+ c-Kit+ (LSK) | Treatment:[13C, 15N] AAs were incorporated for 24 h and subsequently washed out for 0 h | Batch:1b | Note:[13C, 15N] AA levels and [13C]TCA substrate levels in different time point were calculated for AA catabolism (Fig1.C). RAW_FILE_NAME=lsk-0h-1.cdf SUBJECT_SAMPLE_FACTORS - LSK_0h_2 Source_Name[gating]:Lin– Scal1+ c-Kit+ (LSK) | Treatment:[13C, 15N] AAs were incorporated for 24 h and subsequently washed out for 0 h | Batch:1b | Note:[13C, 15N] AA levels and [13C]TCA substrate levels in different time point were calculated for AA catabolism (Fig1.C). RAW_FILE_NAME=lsk-0h-2.cdf SUBJECT_SAMPLE_FACTORS - LSK_0h_3 Source_Name[gating]:Lin– Scal1+ c-Kit+ (LSK) | Treatment:[13C, 15N] AAs were incorporated for 24 h and subsequently washed out for 0 h | Batch:1b | Note:[13C, 15N] AA levels and [13C]TCA substrate levels in different time point were calculated for AA catabolism (Fig1.C). RAW_FILE_NAME=lsk-0h-3.cdf SUBJECT_SAMPLE_FACTORS - LSK_12h_1 Source_Name[gating]:Lin– Scal1+ c-Kit+ (LSK) | Treatment:[13C, 15N] AAs were incorporated for 24 h and subsequently washed out for 12 h | Batch:1b | Note:[13C, 15N] AA levels and [13C]TCA substrate levels in different time point were calculated for AA catabolism (Fig1.C). RAW_FILE_NAME=lsk-12h-1.cdf SUBJECT_SAMPLE_FACTORS - LSK_12h_2 Source_Name[gating]:Lin– Scal1+ c-Kit+ (LSK) | Treatment:[13C, 15N] AAs were incorporated for 24 h and subsequently washed out for 12 h | Batch:1b | Note:[13C, 15N] AA levels and [13C]TCA substrate levels in different time point were calculated for AA catabolism (Fig1.C). RAW_FILE_NAME=lsk-12h-2.cdf SUBJECT_SAMPLE_FACTORS - LSK_12h_3 Source_Name[gating]:Lin– Scal1+ c-Kit+ (LSK) | Treatment:[13C, 15N] AAs were incorporated for 24 h and subsequently washed out for 12 h | Batch:1b | Note:[13C, 15N] AA levels and [13C]TCA substrate levels in different time point were calculated for AA catabolism (Fig1.C). RAW_FILE_NAME=lsk-12h-3.cdf SUBJECT_SAMPLE_FACTORS - LSK_6h_1 Source_Name[gating]:Lin– Scal1+ c-Kit+ (LSK) | Treatment:[13C, 15N] AAs were incorporated for 24 h and subsequently washed out for 6 h | Batch:1b | Note:[13C, 15N] AA levels and [13C]TCA substrate levels in different time point were calculated for AA catabolism (Fig1.C). RAW_FILE_NAME=lsk-6h-1.cdf SUBJECT_SAMPLE_FACTORS - LSK_6h_2 Source_Name[gating]:Lin– Scal1+ c-Kit+ (LSK) | Treatment:[13C, 15N] AAs were incorporated for 24 h and subsequently washed out for 6 h | Batch:1b | Note:[13C, 15N] AA levels and [13C]TCA substrate levels in different time point were calculated for AA catabolism (Fig1.C). RAW_FILE_NAME=lsk-6h-2.cdf SUBJECT_SAMPLE_FACTORS - LSK_6h_3 Source_Name[gating]:Lin– Scal1+ c-Kit+ (LSK) | Treatment:[13C, 15N] AAs were incorporated for 24 h and subsequently washed out for 6 h | Batch:1b | Note:[13C, 15N] AA levels and [13C]TCA substrate levels in different time point were calculated for AA catabolism (Fig1.C). RAW_FILE_NAME=lsk-6h-3.cdf SUBJECT_SAMPLE_FACTORS - Lin+_1 Source_Name[gating]:Mature hematopoietic cells (Lin+ cells) | Treatment:[13C, 15N] AAs were incorporated for 1 h | Batch:1a | Note:[13C, 15N] AAs were used for AA uptake (Fig1.B); unlabeled AAs were used for total AA (Fig1.A) and TCA substrate levels (Fig1.D). RAW_FILE_NAME=linp-1.cdf SUBJECT_SAMPLE_FACTORS - Lin+_2 Source_Name[gating]:Mature hematopoietic cells (Lin+ cells) | Treatment:[13C, 15N] AAs were incorporated for 1 h | Batch:1a | Note:[13C, 15N] AAs were used for AA uptake (Fig1.B); unlabeled AAs were used for total AA (Fig1.A) and TCA substrate levels (Fig1.D). RAW_FILE_NAME=linp-2.cdf SUBJECT_SAMPLE_FACTORS - Lin+_3 Source_Name[gating]:Mature hematopoietic cells (Lin+ cells) | Treatment:[13C, 15N] AAs were incorporated for 1 h | Batch:1a | Note:[13C, 15N] AAs were used for AA uptake (Fig1.B); unlabeled AAs were used for total AA (Fig1.A) and TCA substrate levels (Fig1.D). RAW_FILE_NAME=linp-3.cdf SUBJECT_SAMPLE_FACTORS - Lin+_0h_1 Source_Name[gating]:Mature hematopoietic cells (Lin+ cells) | Treatment:[13C, 15N] AAs were incorporated for 24 h and subsequently washed out for 0 h | Batch:1b | Note:[13C, 15N] AA levels and [13C]TCA substrate levels in different time point were calculated for AA catabolism (Fig1.C). RAW_FILE_NAME=linp-0h-1.cdf SUBJECT_SAMPLE_FACTORS - Lin+_0h_2 Source_Name[gating]:Mature hematopoietic cells (Lin+ cells) | Treatment:[13C, 15N] AAs were incorporated for 24 h and subsequently washed out for 0 h | Batch:1b | Note:[13C, 15N] AA levels and [13C]TCA substrate levels in different time point were calculated for AA catabolism (Fig1.C). RAW_FILE_NAME=linp-0h-2.cdf SUBJECT_SAMPLE_FACTORS - Lin+_0h_3 Source_Name[gating]:Mature hematopoietic cells (Lin+ cells) | Treatment:[13C, 15N] AAs were incorporated for 24 h and subsequently washed out for 0 h | Batch:1b | Note:[13C, 15N] AA levels and [13C]TCA substrate levels in different time point were calculated for AA catabolism (Fig1.C). RAW_FILE_NAME=linp-0h-3.cdf SUBJECT_SAMPLE_FACTORS - Lin+_12h_1 Source_Name[gating]:Mature hematopoietic cells (Lin+ cells) | Treatment:[13C, 15N] AAs were incorporated for 24 h and subsequently washed out for 12 h | Batch:1b | Note:[13C, 15N] AA levels and [13C]TCA substrate levels in different time point were calculated for AA catabolism (Fig1.C). RAW_FILE_NAME=linp-12h-1.cdf SUBJECT_SAMPLE_FACTORS - Lin+_12h_2 Source_Name[gating]:Mature hematopoietic cells (Lin+ cells) | Treatment:[13C, 15N] AAs were incorporated for 24 h and subsequently washed out for 12 h | Batch:1b | Note:[13C, 15N] AA levels and [13C]TCA substrate levels in different time point were calculated for AA catabolism (Fig1.C). RAW_FILE_NAME=linp-12h-2.cdf SUBJECT_SAMPLE_FACTORS - Lin+_12h_3 Source_Name[gating]:Mature hematopoietic cells (Lin+ cells) | Treatment:[13C, 15N] AAs were incorporated for 24 h and subsequently washed out for 12 h | Batch:1b | Note:[13C, 15N] AA levels and [13C]TCA substrate levels in different time point were calculated for AA catabolism (Fig1.C). RAW_FILE_NAME=linp-12h-3.cdf SUBJECT_SAMPLE_FACTORS - Lin+_6h_1 Source_Name[gating]:Mature hematopoietic cells (Lin+ cells) | Treatment:[13C, 15N] AAs were incorporated for 24 h and subsequently washed out for 6 h | Batch:1b | Note:[13C, 15N] AA levels and [13C]TCA substrate levels in different time point were calculated for AA catabolism (Fig1.C). RAW_FILE_NAME=linp-6h-1.cdf SUBJECT_SAMPLE_FACTORS - Lin+_6h_2 Source_Name[gating]:Mature hematopoietic cells (Lin+ cells) | Treatment:[13C, 15N] AAs were incorporated for 24 h and subsequently washed out for 6 h | Batch:1b | Note:[13C, 15N] AA levels and [13C]TCA substrate levels in different time point were calculated for AA catabolism (Fig1.C). RAW_FILE_NAME=linp-6h-2.cdf SUBJECT_SAMPLE_FACTORS - Lin+_6h_3 Source_Name[gating]:Mature hematopoietic cells (Lin+ cells) | Treatment:[13C, 15N] AAs were incorporated for 24 h and subsequently washed out for 6 h | Batch:1b | Note:[13C, 15N] AA levels and [13C]TCA substrate levels in different time point were calculated for AA catabolism (Fig1.C). RAW_FILE_NAME=linp-6h-3.cdf #COLLECTION CO:COLLECTION_SUMMARY 5×10^5 indicated cells were incubated with 100 µM stable [13C,15N] amino acids CO:COLLECTION_SUMMARY (MSK-A2-US-1.2, Cambridge Isotope Laboratories) for indicated time and CO:COLLECTION_SUMMARY centrifuged at 500 g for 5 min at 4 °C. The pelleted cells were extracted in CO:COLLECTION_SUMMARY 500 µl ice-cold Acetonitrile: Isopropyl Alcohol: water (3:3:2 v/v/v) and CO:COLLECTION_SUMMARY aliquoted as three technical replicates. The extracts were vortexed for 5 min at CO:COLLECTION_SUMMARY 4 °C and centrifuged at 14,000 g for 2 min at 4 °C. The supernatants were CO:COLLECTION_SUMMARY dried by vacuum spin for subsequent derivatization and stored at -20 ℃. CO:SAMPLE_TYPE Bone marrow #TREATMENT TR:TREATMENT_SUMMARY Germ-free C57BL/6J mice were intraperitoneally injected with 10 mg/kg 5FU TR:TREATMENT_SUMMARY (F6627-5G, Sigma-Aldrich) for 14 days, or fed with vehicle or NR (400 mg/kg per TR:TREATMENT_SUMMARY day) (1341-23-7, ziyi-reagent) for consecutive 8 weeks (long-term) or 1 week TR:TREATMENT_SUMMARY (short-term). Hematopoietic cells were stained and sorted from treated mice. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY To get the corresponding derivatives, the dried aliquots were incubated with 20 SP:SAMPLEPREP_SUMMARY µL 2% (w/v) methoxyamine hydrochloride (226904, Sigma-Aldrich) in pyridine for SP:SAMPLEPREP_SUMMARY 60 min at 37 °C, and silylated by 30 µL of SP:SAMPLEPREP_SUMMARY N-Methyl-N-(tert-butyldimethylsilyl) trifluoroacetamide with 1% SP:SAMPLEPREP_SUMMARY tert-Butyldimethylchlorosilane (TBDMS, 18162-48-6, Regis Technologies) for 30 SP:SAMPLEPREP_SUMMARY min at 45 °C. The corresponding derivatives were analyzed by GC-MS using the SP:SAMPLEPREP_SUMMARY Trace 1310 gas chromatograph (Thermo Fisher) with the DB-35ms column (Agilent SP:SAMPLEPREP_SUMMARY Technologies) connected to the Q ExactiveTM GC OrbitrapTM GC-MS/MS system SP:SAMPLEPREP_SUMMARY (Thermo Fisher). #CHROMATOGRAPHY CH:INSTRUMENT_NAME TRACE 1310 CH:COLUMN_NAME DB-35ms CH:CHROMATOGRAPHY_TYPE GC #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Thermo Q Exactive Orbitrap MS:INSTRUMENT_TYPE Orbitrap MS:MS_TYPE EI MS:MS_COMMENTS GC-MS data was acquired on Q-Exactive Orbitrap mass spectrometer (Thermo Fisher) MS:MS_COMMENTS coupled with Gas Chromatograph system TRACE 1310 (Thermo Fisher). MS:MS_COMMENTS Chromatographic separation was performed on a DB-35ms column module (30 m length MS:MS_COMMENTS x 0.25 mm internal diameter, Agilent Technologies). The column temperature was MS:MS_COMMENTS programmed with an initial temperature of 50 °C for 2 min, then ramped at MS:MS_COMMENTS 10°C/min to 325 °C, and maintained for 5 min. The mass range was set as 50-600 MS:MS_COMMENTS m/z, and the resolution was 60,000. The ion source temperature was 300°C with MS:MS_COMMENTS the transfer line temperature of 250°C, and the electron energy was 70 eV with MS:MS_COMMENTS EI source. MS:ION_MODE POSITIVE #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS Peak Area MS_METABOLITE_DATA_START Samples Lin–_1 Lin–_2 Lin–_3 Lin–_0h_1 Lin–_0h_2 Lin–_0h_3 Lin–_12h_1 Lin–_12h_2 Lin–_12h_3 Lin–_6h_1 Lin–_6h_2 Lin–_6h_3 HSC_1 HSC_2 HSC_3 HSC_Ctrl_0h_1 HSC_Ctrl_0h_2 HSC_Ctrl_0h_3 HSC_Ctrl_12h_1 HSC_Ctrl_12h_2 HSC_Ctrl_12h_3 HSC_Ctrl_6h_1 HSC_Ctrl_6h_2 HSC_Ctrl_6h_3 LSK_1 LSK_2 LSK_3 LSK_0h_1 LSK_0h_2 LSK_0h_3 LSK_12h_1 LSK_12h_2 LSK_12h_3 LSK_6h_1 LSK_6h_2 LSK_6h_3 Lin+_1 Lin+_2 Lin+_3 Lin+_0h_1 Lin+_0h_2 Lin+_0h_3 Lin+_12h_1 Lin+_12h_2 Lin+_12h_3 Lin+_6h_1 Lin+_6h_2 Lin+_6h_3 Factors Source_Name[gating]:Lineage-negative (Lin–) | Treatment:[13C, 15N] AAs were incorporated for 1 h | Batch:1a | Note:[13C, 15N] AAs were used for AA uptake (Fig1.B); unlabeled AAs were used for total AA (Fig1.A) and TCA substrate levels (Fig1.D). Source_Name[gating]:Lineage-negative (Lin–) | Treatment:[13C, 15N] AAs were incorporated for 1 h | Batch:1a | Note:[13C, 15N] AAs were used for AA uptake (Fig1.B); unlabeled AAs were used for total AA (Fig1.A) and TCA substrate levels (Fig1.D). Source_Name[gating]:Lineage-negative (Lin–) | Treatment:[13C, 15N] AAs were incorporated for 1 h | Batch:1a | Note:[13C, 15N] AAs were used for AA uptake (Fig1.B); unlabeled AAs were used for total AA (Fig1.A) and TCA substrate levels (Fig1.D). Source_Name[gating]:Lineage-negative (Lin–) | Treatment:[13C, 15N] AAs were incorporated for 24 h and subsequently washed out for 0 h | Batch:1b | Note:[13C, 15N] AA levels and [13C]TCA substrate levels in different time point were calculated for AA catabolism (Fig1.C). Source_Name[gating]:Lineage-negative (Lin–) | Treatment:[13C, 15N] AAs were incorporated for 24 h and subsequently washed out for 0 h | Batch:1b | Note:[13C, 15N] AA levels and [13C]TCA substrate levels in different time point were calculated for AA catabolism (Fig1.C). Source_Name[gating]:Lineage-negative (Lin–) | Treatment:[13C, 15N] AAs were incorporated for 24 h and subsequently washed out for 0 h | Batch:1b | Note:[13C, 15N] AA levels and [13C]TCA substrate levels in different time point were calculated for AA catabolism (Fig1.C). Source_Name[gating]:Lineage-negative (Lin–) | Treatment:[13C, 15N] AAs were incorporated for 24 h and subsequently washed out for 12 h | Batch:1b | Note:[13C, 15N] AA levels and [13C]TCA substrate levels in different time point were calculated for AA catabolism (Fig1.C). Source_Name[gating]:Lineage-negative (Lin–) | Treatment:[13C, 15N] AAs were incorporated for 24 h and subsequently washed out for 12 h | Batch:1b | Note:[13C, 15N] AA levels and [13C]TCA substrate levels in different time point were calculated for AA catabolism (Fig1.C). Source_Name[gating]:Lineage-negative (Lin–) | Treatment:[13C, 15N] AAs were incorporated for 24 h and subsequently washed out for 12 h | Batch:1b | Note:[13C, 15N] AA levels and [13C]TCA substrate levels in different time point were calculated for AA catabolism (Fig1.C). Source_Name[gating]:Lineage-negative (Lin–) | Treatment:[13C, 15N] AAs were incorporated for 24 h and subsequently washed out for 6 h | Batch:1b | Note:[13C, 15N] AA levels and [13C]TCA substrate levels in different time point were calculated for AA catabolism (Fig1.C). Source_Name[gating]:Lineage-negative (Lin–) | Treatment:[13C, 15N] AAs were incorporated for 24 h and subsequently washed out for 6 h | Batch:1b | Note:[13C, 15N] AA levels and [13C]TCA substrate levels in different time point were calculated for AA catabolism (Fig1.C). Source_Name[gating]:Lineage-negative (Lin–) | Treatment:[13C, 15N] AAs were incorporated for 24 h and subsequently washed out for 6 h | Batch:1b | Note:[13C, 15N] AA levels and [13C]TCA substrate levels in different time point were calculated for AA catabolism (Fig1.C). Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:[13C, 15N] AAs were incorporated for 1 h | Batch:1a | Note:[13C, 15N] AAs were used for AA uptake (Fig1.B); unlabeled AAs were used for total AA (Fig1.A) and TCA substrate levels (Fig1.D). Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:[13C, 15N] AAs were incorporated for 1 h | Batch:1a | Note:[13C, 15N] AAs were used for AA uptake (Fig1.B); unlabeled AAs were used for total AA (Fig1.A) and TCA substrate levels (Fig1.D). Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:[13C, 15N] AAs were incorporated for 1 h | Batch:1a | Note:[13C, 15N] AAs were used for AA uptake (Fig1.B); unlabeled AAs were used for total AA (Fig1.A) and TCA substrate levels (Fig1.D). Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:HSC Basal, [13C, 15N] AAs were incorporated for 24 h and subsequently washed out for 0 h | Batch:1b | Note:[13C, 15N] AA levels and [13C]TCA substrate levels in different time point were calculated for AA catabolism (Fig1.C, Fig2.D). Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:HSC Basal, [13C, 15N] AAs were incorporated for 24 h and subsequently washed out for 0 h | Batch:1b | Note:[13C, 15N] AA levels and [13C]TCA substrate levels in different time point were calculated for AA catabolism (Fig1.C, Fig2.D). Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:HSC Basal, [13C, 15N] AAs were incorporated for 24 h and subsequently washed out for 0 h | Batch:1b | Note:[13C, 15N] AA levels and [13C]TCA substrate levels in different time point were calculated for AA catabolism (Fig1.C, Fig2.D). Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:HSC Basal, [13C, 15N] AAs were incorporated for 24 h and subsequently washed out for 12 h | Batch:1b | Note:[13C, 15N] AA levels and [13C]TCA substrate levels in different time point were calculated for AA catabolism (Fig1.C, Fig2.D). Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:HSC Basal, [13C, 15N] AAs were incorporated for 24 h and subsequently washed out for 12 h | Batch:1b | Note:[13C, 15N] AA levels and [13C]TCA substrate levels in different time point were calculated for AA catabolism (Fig1.C, Fig2.D). Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:HSC Basal, [13C, 15N] AAs were incorporated for 24 h and subsequently washed out for 12 h | Batch:1b | Note:[13C, 15N] AA levels and [13C]TCA substrate levels in different time point were calculated for AA catabolism (Fig1.C, Fig2.D). Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:HSC Basal, [13C, 15N] AAs were incorporated for 24 h and subsequently washed out for 6 h | Batch:1b | Note:[13C, 15N] AA levels and [13C]TCA substrate levels in different time point were calculated for AA catabolism (Fig1.C, Fig2.D). Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:HSC Basal, [13C, 15N] AAs were incorporated for 24 h and subsequently washed out for 6 h | Batch:1b | Note:[13C, 15N] AA levels and [13C]TCA substrate levels in different time point were calculated for AA catabolism (Fig1.C, Fig2.D). Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:HSC Basal, [13C, 15N] AAs were incorporated for 24 h and subsequently washed out for 6 h | Batch:1b | Note:[13C, 15N] AA levels and [13C]TCA substrate levels in different time point were calculated for AA catabolism (Fig1.C, Fig2.D). Source_Name[gating]:Lin– Scal1+ c-Kit+ (LSK) | Treatment:[13C, 15N] AAs were incorporated for 1 h | Batch:1a | Note:[13C, 15N] AAs were used for AA uptake (Fig1.B); unlabeled AAs were used for total AA (Fig1.A) and TCA substrate levels (Fig1.D). Source_Name[gating]:Lin– Scal1+ c-Kit+ (LSK) | Treatment:[13C, 15N] AAs were incorporated for 1 h | Batch:1a | Note:[13C, 15N] AAs were used for AA uptake (Fig1.B); unlabeled AAs were used for total AA (Fig1.A) and TCA substrate levels (Fig1.D). Source_Name[gating]:Lin– Scal1+ c-Kit+ (LSK) | Treatment:[13C, 15N] AAs were incorporated for 1 h | Batch:1a | Note:[13C, 15N] AAs were used for AA uptake (Fig1.B); unlabeled AAs were used for total AA (Fig1.A) and TCA substrate levels (Fig1.D). Source_Name[gating]:Lin– Scal1+ c-Kit+ (LSK) | Treatment:[13C, 15N] AAs were incorporated for 24 h and subsequently washed out for 0 h | Batch:1b | Note:[13C, 15N] AA levels and [13C]TCA substrate levels in different time point were calculated for AA catabolism (Fig1.C). Source_Name[gating]:Lin– Scal1+ c-Kit+ (LSK) | Treatment:[13C, 15N] AAs were incorporated for 24 h and subsequently washed out for 0 h | Batch:1b | Note:[13C, 15N] AA levels and [13C]TCA substrate levels in different time point were calculated for AA catabolism (Fig1.C). Source_Name[gating]:Lin– Scal1+ c-Kit+ (LSK) | Treatment:[13C, 15N] AAs were incorporated for 24 h and subsequently washed out for 0 h | Batch:1b | Note:[13C, 15N] AA levels and [13C]TCA substrate levels in different time point were calculated for AA catabolism (Fig1.C). Source_Name[gating]:Lin– Scal1+ c-Kit+ (LSK) | Treatment:[13C, 15N] AAs were incorporated for 24 h and subsequently washed out for 12 h | Batch:1b | Note:[13C, 15N] AA levels and [13C]TCA substrate levels in different time point were calculated for AA catabolism (Fig1.C). Source_Name[gating]:Lin– Scal1+ c-Kit+ (LSK) | Treatment:[13C, 15N] AAs were incorporated for 24 h and subsequently washed out for 12 h | Batch:1b | Note:[13C, 15N] AA levels and [13C]TCA substrate levels in different time point were calculated for AA catabolism (Fig1.C). Source_Name[gating]:Lin– Scal1+ c-Kit+ (LSK) | Treatment:[13C, 15N] AAs were incorporated for 24 h and subsequently washed out for 12 h | Batch:1b | Note:[13C, 15N] AA levels and [13C]TCA substrate levels in different time point were calculated for AA catabolism (Fig1.C). Source_Name[gating]:Lin– Scal1+ c-Kit+ (LSK) | Treatment:[13C, 15N] AAs were incorporated for 24 h and subsequently washed out for 6 h | Batch:1b | Note:[13C, 15N] AA levels and [13C]TCA substrate levels in different time point were calculated for AA catabolism (Fig1.C). Source_Name[gating]:Lin– Scal1+ c-Kit+ (LSK) | Treatment:[13C, 15N] AAs were incorporated for 24 h and subsequently washed out for 6 h | Batch:1b | Note:[13C, 15N] AA levels and [13C]TCA substrate levels in different time point were calculated for AA catabolism (Fig1.C). Source_Name[gating]:Lin– Scal1+ c-Kit+ (LSK) | Treatment:[13C, 15N] AAs were incorporated for 24 h and subsequently washed out for 6 h | Batch:1b | Note:[13C, 15N] AA levels and [13C]TCA substrate levels in different time point were calculated for AA catabolism (Fig1.C). Source_Name[gating]:Mature hematopoietic cells (Lin+ cells) | Treatment:[13C, 15N] AAs were incorporated for 1 h | Batch:1a | Note:[13C, 15N] AAs were used for AA uptake (Fig1.B); unlabeled AAs were used for total AA (Fig1.A) and TCA substrate levels (Fig1.D). Source_Name[gating]:Mature hematopoietic cells (Lin+ cells) | Treatment:[13C, 15N] AAs were incorporated for 1 h | Batch:1a | Note:[13C, 15N] AAs were used for AA uptake (Fig1.B); unlabeled AAs were used for total AA (Fig1.A) and TCA substrate levels (Fig1.D). Source_Name[gating]:Mature hematopoietic cells (Lin+ cells) | Treatment:[13C, 15N] AAs were incorporated for 1 h | Batch:1a | Note:[13C, 15N] AAs were used for AA uptake (Fig1.B); unlabeled AAs were used for total AA (Fig1.A) and TCA substrate levels (Fig1.D). Source_Name[gating]:Mature hematopoietic cells (Lin+ cells) | Treatment:[13C, 15N] AAs were incorporated for 24 h and subsequently washed out for 0 h | Batch:1b | Note:[13C, 15N] AA levels and [13C]TCA substrate levels in different time point were calculated for AA catabolism (Fig1.C). Source_Name[gating]:Mature hematopoietic cells (Lin+ cells) | Treatment:[13C, 15N] AAs were incorporated for 24 h and subsequently washed out for 0 h | Batch:1b | Note:[13C, 15N] AA levels and [13C]TCA substrate levels in different time point were calculated for AA catabolism (Fig1.C). Source_Name[gating]:Mature hematopoietic cells (Lin+ cells) | Treatment:[13C, 15N] AAs were incorporated for 24 h and subsequently washed out for 0 h | Batch:1b | Note:[13C, 15N] AA levels and [13C]TCA substrate levels in different time point were calculated for AA catabolism (Fig1.C). Source_Name[gating]:Mature hematopoietic cells (Lin+ cells) | Treatment:[13C, 15N] AAs were incorporated for 24 h and subsequently washed out for 12 h | Batch:1b | Note:[13C, 15N] AA levels and [13C]TCA substrate levels in different time point were calculated for AA catabolism (Fig1.C). Source_Name[gating]:Mature hematopoietic cells (Lin+ cells) | Treatment:[13C, 15N] AAs were incorporated for 24 h and subsequently washed out for 12 h | Batch:1b | Note:[13C, 15N] AA levels and [13C]TCA substrate levels in different time point were calculated for AA catabolism (Fig1.C). Source_Name[gating]:Mature hematopoietic cells (Lin+ cells) | Treatment:[13C, 15N] AAs were incorporated for 24 h and subsequently washed out for 12 h | Batch:1b | Note:[13C, 15N] AA levels and [13C]TCA substrate levels in different time point were calculated for AA catabolism (Fig1.C). Source_Name[gating]:Mature hematopoietic cells (Lin+ cells) | Treatment:[13C, 15N] AAs were incorporated for 24 h and subsequently washed out for 6 h | Batch:1b | Note:[13C, 15N] AA levels and [13C]TCA substrate levels in different time point were calculated for AA catabolism (Fig1.C). Source_Name[gating]:Mature hematopoietic cells (Lin+ cells) | Treatment:[13C, 15N] AAs were incorporated for 24 h and subsequently washed out for 6 h | Batch:1b | Note:[13C, 15N] AA levels and [13C]TCA substrate levels in different time point were calculated for AA catabolism (Fig1.C). Source_Name[gating]:Mature hematopoietic cells (Lin+ cells) | Treatment:[13C, 15N] AAs were incorporated for 24 h and subsequently washed out for 6 h | Batch:1b | Note:[13C, 15N] AA levels and [13C]TCA substrate levels in different time point were calculated for AA catabolism (Fig1.C). [13C_ 15N]Alanine_ 2TBDMS 55300.0000 47500.0000 54000.0000 5530000.0000 4750000.0000 5400000.0000 4740000.0000 3940000.0000 4860000.0000 5130000.0000 4520000.0000 5020000.0000 65000.0000 72000.0000 62900.0000 6500000.0000 7200000.0000 6290000.0000 4760000.0000 5250000.0000 4820000.0000 5550000.0000 6110000.0000 5630000.0000 51900.0000 58400.0000 57100.0000 5190000.0000 5840000.0000 5710000.0000 4960000.0000 4350000.0000 4920000.0000 5030000.0000 5230000.0000 5230000.0000 53800.0000 46200.0000 45600.0000 5380000.0000 4620000.0000 4560000.0000 4190000.0000 4280000.0000 4180000.0000 5120000.0000 4200000.0000 4330000.0000 [13C_ 15N]Aspartic acid_ 3TBDMS 72100.0000 72600.0000 79100.0000 7210000.0000 7260000.0000 7910000.0000 6930000.0000 7030000.0000 7240000.0000 7120000.0000 7130000.0000 7420000.0000 86900.0000 91200.0000 89000.0000 8690000.0000 9120000.0000 8900000.0000 2590000.0000 4080000.0000 3610000.0000 4330000.0000 6320000.0000 5630000.0000 81300.0000 78200.0000 79300.0000 8130000.0000 7820000.0000 7930000.0000 7930000.0000 7160000.0000 7290000.0000 8030000.0000 7430000.0000 7510000.0000 74300.0000 71300.0000 72100.0000 7430000.0000 7130000.0000 7210000.0000 7300000.0000 6760000.0000 6780000.0000 7400000.0000 6970000.0000 7010000.0000 [13C_ 15N]Glutamic acid_ 3TBDMS 13100.0000 12600.0000 13800.0000 1310000.0000 1260000.0000 1380000.0000 1020000.0000 1100000.0000 1080000.0000 1240000.0000 1200000.0000 1240000.0000 30900.0000 25900.0000 26200.0000 3090000.0000 2590000.0000 2620000.0000 440000.0000 657000.0000 588000.0000 1750000.0000 1500000.0000 1650000.0000 18000.0000 15900.0000 14600.0000 1800000.0000 1590000.0000 1460000.0000 1140000.0000 1120000.0000 1080000.0000 1350000.0000 1330000.0000 1600000.0000 12800.0000 12000.0000 11300.0000 1280000.0000 1200000.0000 1130000.0000 982000.0000 1050000.0000 954000.0000 1040000.0000 1130000.0000 1020000.0000 [13C_ 15N]Glycine_ 2TBDMS 47700.0000 63000.0000 45000.0000 3830000.0000 3930000.0000 3070000.0000 3250000.0000 3540000.0000 2660000.0000 3530000.0000 3740000.0000 4060000.0000 63200.0000 53000.0000 63200.0000 6490000.0000 5780000.0000 4740000.0000 3690000.0000 2190000.0000 2590000.0000 3180000.0000 5300000.0000 3670000.0000 38100.0000 39900.0000 34700.0000 4560000.0000 5870000.0000 4060000.0000 3140000.0000 3500000.0000 4630000.0000 4060000.0000 5260000.0000 3360000.0000 49500.0000 39800.0000 49500.0000 3470000.0000 3610000.0000 2720000.0000 3290000.0000 3620000.0000 3170000.0000 3640000.0000 3180000.0000 3440000.0000 [13C_ 15N]Histidine_ 3TBDMS 21800.0000 23000.0000 32700.0000 2180000.0000 2300000.0000 3270000.0000 1760000.0000 2170000.0000 3080000.0000 2030000.0000 2210000.0000 3120000.0000 30600.0000 34800.0000 35000.0000 3060000.0000 3480000.0000 3500000.0000 1120000.0000 936000.0000 853000.0000 2150000.0000 2230000.0000 2530000.0000 29600.0000 21900.0000 24800.0000 2960000.0000 2190000.0000 2480000.0000 2870000.0000 1910000.0000 2110000.0000 2910000.0000 2010000.0000 2230000.0000 28200.0000 27100.0000 32900.0000 2820000.0000 2710000.0000 3290000.0000 2690000.0000 2530000.0000 3020000.0000 2730000.0000 2630000.0000 3130000.0000 [13C_ 15N]Isoleucine_ 2TBDMS 93200.0000 93900.0000 86500.0000 9320000.0000 9390000.0000 8650000.0000 7020000.0000 6850000.0000 6870000.0000 8130000.0000 8030000.0000 8230000.0000 94500.0000 132000.0000 115000.0000 9450000.0000 13200000.0000 11500000.0000 3430000.0000 3140000.0000 4050000.0000 6520000.0000 6120000.0000 7130000.0000 96800.0000 97000.0000 90600.0000 9680000.0000 9700000.0000 9060000.0000 6450000.0000 7110000.0000 7110000.0000 8130000.0000 8430000.0000 8210000.0000 86700.0000 83400.0000 80300.0000 8670000.0000 8340000.0000 8030000.0000 6210000.0000 6670000.0000 6740000.0000 8410000.0000 8010000.0000 7630000.0000 [13C_ 15N]Leucine_ 2TBDMS 90500.0000 93400.0000 97200.0000 9050000.0000 9340000.0000 9720000.0000 8770000.0000 8380000.0000 8720000.0000 8830000.0000 9130000.0000 9320000.0000 103000.0000 149000.0000 129000.0000 10300000.0000 14900000.0000 12900000.0000 4660000.0000 4060000.0000 4370000.0000 8610000.0000 8120000.0000 8310000.0000 90700.0000 98400.0000 90200.0000 9070000.0000 9840000.0000 9020000.0000 8790000.0000 9470000.0000 8360000.0000 8930000.0000 9530000.0000 8630000.0000 89500.0000 93700.0000 87200.0000 8950000.0000 9370000.0000 8720000.0000 8050000.0000 8130000.0000 8510000.0000 8420000.0000 9120000.0000 8630000.0000 [13C_ 15N]Lysine_ 3TBDMS 45500.0000 49000.0000 58700.0000 4550000.0000 4900000.0000 5870000.0000 4180000.0000 4400000.0000 4280000.0000 4240000.0000 4630000.0000 4750000.0000 56300.0000 78400.0000 62200.0000 5630000.0000 7840000.0000 6220000.0000 2980000.0000 3580000.0000 3040000.0000 5650000.0000 4520000.0000 5970000.0000 44500.0000 51100.0000 53300.0000 4450000.0000 5110000.0000 5330000.0000 3910000.0000 4550000.0000 4620000.0000 4120000.0000 4830000.0000 4960000.0000 42000.0000 55400.0000 47400.0000 4200000.0000 5540000.0000 4740000.0000 4130000.0000 4320000.0000 4270000.0000 4120000.0000 4730000.0000 4630000.0000 [13C_ 15N]Methionine_ 2TBDMS 83100.0000 89600.0000 80300.0000 8310000.0000 8960000.0000 8030000.0000 8280000.0000 8210000.0000 7560000.0000 8300000.0000 8410000.0000 7830000.0000 96600.0000 128000.0000 155000.0000 9660000.0000 12800000.0000 15500000.0000 7580000.0000 6760000.0000 7070000.0000 8510000.0000 8630000.0000 8510000.0000 112000.0000 94000.0000 93900.0000 11200000.0000 9400000.0000 9390000.0000 9280000.0000 8330000.0000 8170000.0000 9860000.0000 9130000.0000 9120000.0000 83900.0000 81600.0000 79600.0000 8390000.0000 8160000.0000 7960000.0000 8090000.0000 8090000.0000 7380000.0000 8100000.0000 8100000.0000 7430000.0000 [13C_ 15N]Phenylalanine_ 2TBDMS 94000.0000 90600.0000 114000.0000 9400000.0000 9060000.0000 11400000.0000 8020000.0000 8610000.0000 8140000.0000 8230000.0000 8810000.0000 8210000.0000 114000.0000 159000.0000 112000.0000 11400000.0000 15900000.0000 11200000.0000 5630000.0000 4620000.0000 4530000.0000 8530000.0000 8420000.0000 7510000.0000 111000.0000 129000.0000 110000.0000 11100000.0000 12900000.0000 11000000.0000 7320000.0000 8150000.0000 7540000.0000 8820000.0000 8300000.0000 9130000.0000 82700.0000 81400.0000 87700.0000 8270000.0000 8140000.0000 8770000.0000 8010000.0000 7750000.0000 8070000.0000 8100000.0000 7960000.0000 8350000.0000 [13C_ 15N]Proline_ 2TBDMS 47500.0000 41800.0000 51000.0000 4750000.0000 4180000.0000 5100000.0000 3790000.0000 3430000.0000 4230000.0000 4140000.0000 3920000.0000 4630000.0000 75700.0000 87700.0000 75600.0000 7570000.0000 8770000.0000 7560000.0000 2800000.0000 1980000.0000 2880000.0000 6120000.0000 5250000.0000 5150000.0000 51700.0000 49200.0000 57600.0000 5170000.0000 4920000.0000 5760000.0000 3510000.0000 4660000.0000 3850000.0000 4010000.0000 4710000.0000 4140000.0000 47300.0000 50700.0000 47200.0000 4730000.0000 5070000.0000 4720000.0000 4000000.0000 4490000.0000 4150000.0000 4420000.0000 4730000.0000 4510000.0000 [13C_ 15N]Serine_ 3TBDMS 83300.0000 91300.0000 89200.0000 8330000.0000 9130000.0000 8920000.0000 7510000.0000 8420000.0000 8260000.0000 8130000.0000 8710000.0000 8420000.0000 96000.0000 83400.0000 98600.0000 9600000.0000 8340000.0000 9860000.0000 6160000.0000 6690000.0000 6980000.0000 7330000.0000 7230000.0000 8890000.0000 89700.0000 95100.0000 95300.0000 8970000.0000 9510000.0000 9530000.0000 7460000.0000 8520000.0000 8180000.0000 8130000.0000 9130000.0000 9230000.0000 95800.0000 83100.0000 86200.0000 9580000.0000 8310000.0000 8620000.0000 8860000.0000 8090000.0000 8150000.0000 9130000.0000 8130000.0000 8320000.0000 [13C_ 15N]Threonine_ 3TBDMS 11200.0000 12000.0000 13600.0000 1120000.0000 1200000.0000 1360000.0000 916000.0000 1080000.0000 1010000.0000 1030000.0000 1150000.0000 1200000.0000 16600.0000 15800.0000 20800.0000 1660000.0000 1580000.0000 2080000.0000 795000.0000 889000.0000 885000.0000 1130000.0000 1110000.0000 1250000.0000 13000.0000 12600.0000 13200.0000 1300000.0000 1260000.0000 1320000.0000 1100000.0000 1040000.0000 1020000.0000 1160000.0000 1150000.0000 1210000.0000 12900.0000 12600.0000 11500.0000 1290000.0000 1260000.0000 1150000.0000 1100000.0000 1050000.0000 979000.0000 1140000.0000 1200000.0000 1030000.0000 [13C_ 15N]Tyrosine_ 3TBDMS 351000.0000 280000.0000 312000.0000 35100000.0000 28000000.0000 31200000.0000 26000000.0000 25300000.0000 28100000.0000 31500000.0000 26900000.0000 31000000.0000 474000.0000 540000.0000 445000.0000 47400000.0000 54000000.0000 44500000.0000 16900000.0000 23400000.0000 25200000.0000 35400000.0000 32200000.0000 30400000.0000 336000.0000 331000.0000 293000.0000 33600000.0000 33100000.0000 29300000.0000 28200000.0000 29900000.0000 26800000.0000 31500000.0000 31300000.0000 28300000.0000 344000.0000 335000.0000 294000.0000 34400000.0000 33500000.0000 29400000.0000 31500000.0000 30500000.0000 24100000.0000 32500000.0000 31300000.0000 26600000.0000 [13C_ 15N]Valine_ 2TBDMS 350000.0000 370000.0000 267000.0000 35000000.0000 37000000.0000 26700000.0000 23600000.0000 20900000.0000 22600000.0000 24100000.0000 23500000.0000 24300000.0000 516000.0000 412000.0000 455000.0000 51600000.0000 41200000.0000 45500000.0000 15500000.0000 17200000.0000 14000000.0000 28300000.0000 31400000.0000 25200000.0000 338000.0000 300000.0000 366000.0000 33800000.0000 30000000.0000 36600000.0000 22400000.0000 25100000.0000 22700000.0000 22100000.0000 25300000.0000 24300000.0000 338000.0000 264000.0000 368000.0000 33800000.0000 26400000.0000 36800000.0000 23200000.0000 23700000.0000 23800000.0000 21000000.0000 22500000.0000 24300000.0000 [13C]Alpha_Ketoglutarate 19700.0000 14400.0000 20100.0000 22300.0000 19800.0000 21700.0000 21200.0000 16300.0000 21000.0000 25500.0000 30200.0000 27200.0000 83400.0000 64600.0000 82700.0000 41200.0000 41200.0000 49700.0000 17200.0000 18700.0000 20200.0000 21700.0000 23600.0000 27100.0000 19600.0000 21400.0000 24200.0000 21100.0000 21000.0000 14500.0000 26300.0000 31200.0000 20900.0000 23300.0000 28600.0000 16300.0000 [13C]Citrate 44800.0000 42600.0000 49600.0000 61200.0000 62900.0000 58200.0000 53200.0000 52100.0000 54000.0000 50500.0000 60800.0000 62000.0000 192000.0000 194000.0000 212000.0000 115000.0000 108000.0000 125000.0000 47400.0000 41600.0000 46400.0000 56500.0000 50400.0000 54300.0000 51200.0000 48600.0000 51200.0000 40300.0000 43300.0000 31300.0000 57500.0000 53200.0000 59900.0000 51100.0000 51300.0000 45200.0000 [13C]Fumatate 187000.0000 183000.0000 193000.0000 217000.0000 197000.0000 212000.0000 203000.0000 190000.0000 210000.0000 209000.0000 181000.0000 226000.0000 290000.0000 313000.0000 313000.0000 256000.0000 243000.0000 264000.0000 161000.0000 184000.0000 187000.0000 213000.0000 199000.0000 192000.0000 186000.0000 190000.0000 190000.0000 199000.0000 188000.0000 182000.0000 218000.0000 200000.0000 192000.0000 203000.0000 192000.0000 188000.0000 [13C]Malate 65000.0000 69800.0000 69600.0000 76700.0000 71400.0000 84200.0000 71200.0000 70300.0000 81200.0000 96700.0000 86700.0000 76100.0000 114000.0000 119000.0000 131000.0000 92700.0000 83300.0000 105000.0000 70400.0000 73600.0000 64700.0000 77000.0000 78900.0000 80700.0000 74200.0000 75100.0000 71200.0000 64000.0000 67200.0000 52800.0000 75400.0000 74500.0000 82200.0000 71200.0000 71200.0000 68300.0000 [13C]Succinate 139000.0000 127000.0000 139000.0000 154000.0000 135000.0000 192000.0000 144000.0000 133000.0000 163000.0000 142000.0000 183000.0000 153000.0000 293000.0000 267000.0000 244000.0000 215000.0000 204000.0000 215000.0000 136000.0000 137000.0000 128000.0000 178000.0000 186000.0000 154000.0000 152000.0000 154000.0000 142000.0000 135000.0000 139000.0000 102000.0000 173000.0000 177000.0000 143000.0000 152000.0000 154000.0000 132000.0000 Alanine_ 2TBDMS 5260000.0000 5190000.0000 6140000.0000 1210000.0000 1420000.0000 1680000.0000 3920000.0000 3810000.0000 4970000.0000 6420000.0000 6330000.0000 6210000.0000 Alpha_Ketoglutarate 6210000.0000 6430000.0000 6260000.0000 4190000.0000 4230000.0000 4110000.0000 6020000.0000 5840000.0000 5950000.0000 6140000.0000 6210000.0000 6290000.0000 Aspartic acid_ 3TBDMS 737000000.0000 835000000.0000 735000000.0000 117000000.0000 110000000.0000 127000000.0000 611000000.0000 679000000.0000 591000000.0000 937000000.0000 1120000000.0000 1130000000.0000 Citrate 13400000.0000 13400000.0000 13400000.0000 8850000.0000 7490000.0000 7070000.0000 10000000.0000 12800000.0000 11100000.0000 12300000.0000 12900000.0000 12700000.0000 Fumatate 64400000.0000 64900000.0000 63600000.0000 40800000.0000 42400000.0000 41700000.0000 61200000.0000 62800000.0000 64600000.0000 66600000.0000 63500000.0000 65900000.0000 Glutamic acid_ 3TBDMS 947000000.0000 947000000.0000 847000000.0000 195000000.0000 283000000.0000 241000000.0000 662000000.0000 613000000.0000 735000000.0000 1120000000.0000 1310000000.0000 1020000000.0000 Glutamine_ 3TBDMS 2360000.0000 2190000.0000 2130000.0000 339000.0000 411000.0000 532000.0000 1420000.0000 1180000.0000 1540000.0000 3140000.0000 3010000.0000 3190000.0000 Glycine_ 2TBDMS 1530000000.0000 1480000000.0000 1430000000.0000 546000000.0000 619000000.0000 622000000.0000 1110000000.0000 1290000000.0000 1180000000.0000 1480000000.0000 1530000000.0000 1560000000.0000 Histidine_ 3TBDMS 785000.0000 810000.0000 925000.0000 157000.0000 212000.0000 149000.0000 488000.0000 556000.0000 552000.0000 1130000.0000 1030000.0000 1330000.0000 Hydroxyproline_ (E)-_ 3TBDMS 9500000.0000 9540000.0000 10400000.0000 1730000.0000 2290000.0000 2180000.0000 7750000.0000 6930000.0000 7340000.0000 13200000.0000 14300000.0000 15200000.0000 Isoleucine_ 2TBDMS 21400000.0000 23400000.0000 22100000.0000 5700000.0000 5450000.0000 6060000.0000 16000000.0000 15500000.0000 17600000.0000 25900000.0000 25000000.0000 25500000.0000 L-Cysteine_ 3TBDMS 8730000.0000 8660000.0000 9140000.0000 1360000.0000 932000.0000 1560000.0000 5180000.0000 5420000.0000 5300000.0000 11400000.0000 12500000.0000 14300000.0000 Leucine_ 2TBDMS 89800000.0000 90100000.0000 89300000.0000 26400000.0000 22300000.0000 24600000.0000 67100000.0000 65800000.0000 69600000.0000 94100000.0000 101000000.0000 100000000.0000 Lysine_ 3TBDMS 43700000.0000 53700000.0000 54000000.0000 9020000.0000 12200000.0000 13200000.0000 39300000.0000 39500000.0000 43500000.0000 63800000.0000 62600000.0000 61200000.0000 Malate 19400000.0000 19800000.0000 18400000.0000 11100000.0000 9850000.0000 9250000.0000 16700000.0000 16500000.0000 15600000.0000 20000000.0000 18700000.0000 19800000.0000 Methionine_ 2TBDMS 15200000.0000 16900000.0000 15600000.0000 3440000.0000 3350000.0000 3750000.0000 8110000.0000 8610000.0000 9560000.0000 18300000.0000 19100000.0000 19900000.0000 Phenylalanine_ 2TBDMS 41600000.0000 36000000.0000 37600000.0000 5520000.0000 6450000.0000 7050000.0000 26300000.0000 26500000.0000 27600000.0000 49800000.0000 46800000.0000 41700000.0000 Proline_ 2TBDMS 141000000.0000 131000000.0000 172000000.0000 34100000.0000 30900000.0000 31800000.0000 96900000.0000 87700000.0000 96700000.0000 211000000.0000 228000000.0000 226000000.0000 Serine_ 3TBDMS 316000000.0000 320000000.0000 315000000.0000 63800000.0000 60400000.0000 67500000.0000 242000000.0000 239000000.0000 242000000.0000 325000000.0000 357000000.0000 353000000.0000 Succinate 62300000.0000 65900000.0000 64500000.0000 41200000.0000 43700000.0000 41300000.0000 57000000.0000 57500000.0000 55500000.0000 68500000.0000 63300000.0000 64200000.0000 Threonine_ 3TBDMS 11800000.0000 11900000.0000 12000000.0000 2480000.0000 3190000.0000 2760000.0000 7090000.0000 7470000.0000 7700000.0000 17600000.0000 19100000.0000 18500000.0000 Tyrosine_ 3TBDMS 3130000.0000 3170000.0000 3330000.0000 524000.0000 556000.0000 797000.0000 2470000.0000 3180000.0000 2720000.0000 4510000.0000 4120000.0000 4400000.0000 Valine_ 2TBDMS 54100000.0000 54300000.0000 53700000.0000 15400000.0000 13600000.0000 14000000.0000 38500000.0000 39500000.0000 41300000.0000 57400000.0000 55800000.0000 54100000.0000 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name pubchem_id inchi_key kegg_id other_id other_id_type ri ri_type moverz_quant [13C, 15N]Alanine, 2TBDMS 15.06 [13C, 15N]Aspartic acid, 3TBDMS 21.49 [13C, 15N]Glutamic acid, 3TBDMS 22.54 [13C, 15N]Glycine, 2TBDMS 15.39 [13C, 15N]Histidine, 3TBDMS 25.11 [13C, 15N]Isoleucine, 2TBDMS 17.26 [13C, 15N]Leucine, 2TBDMS 16.91 [13C, 15N]Lysine, 3TBDMS 23.48 [13C, 15N]Methionine, 2TBDMS 19.74 [13C, 15N]Phenylalanine, 2TBDMS 20.92 [13C, 15N]Proline, 2TBDMS 20.49 [13C, 15N]Serine, 3TBDMS 19.94 [13C, 15N]Threonine, 3TBDMS 20.22 [13C, 15N]Tyrosine, 3TBDMS 25.51 [13C, 15N]Valine, 2TBDMS 16.46 [13C]Alpha_Ketoglutarate 21.03 [13C]Citrate 26.51 [13C]Fumatate 18.77 [13C]Malate 22.05 [13C]Succinate 18.37 Alanine, 2TBDMS 15.06 Alpha_Ketoglutarate 21.02 Aspartic acid, 3TBDMS 21.48 Citrate 26.51 Fumatate 18.76 Glutamic acid, 3TBDMS 22.53 Glutamine, 3TBDMS 23.81 Glycine, 2TBDMS 15.38 Histidine, 3TBDMS 25.10 Hydroxyproline, (E)-, 3TBDMS 21.77 Isoleucine, 2TBDMS 17.25 L-Cysteine, 3TBDMS 21.94 Leucine, 2TBDMS 16.90 Lysine, 3TBDMS 23.47 Malate 22.04 Methionine, 2TBDMS 19.73 Phenylalanine, 2TBDMS 20.91 Proline, 2TBDMS 20.48 Serine, 3TBDMS 19.93 Succinate 18.37 Threonine, 3TBDMS 20.21 Tyrosine, 3TBDMS 25.50 Valine, 2TBDMS 16.45 METABOLITES_END #END