#METABOLOMICS WORKBENCH zhaom_20220406_223930 DATATRACK_ID:3179 STUDY_ID:ST002125 ANALYSIS_ID:AN003478 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 (Part2) 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 - BM_supernatant_5FU_1 Source_Name[gating]:Bone marrow supernatant | Treatment:5 FU 2 weeks | Batch:1c | Note:Unlabeled AAs were used for total AA (FigS2.D) RAW_FILE_NAME=bms-fu-1.cdf SUBJECT_SAMPLE_FACTORS - BM_supernatant_5FU_2 Source_Name[gating]:Bone marrow supernatant | Treatment:5 FU 2 weeks | Batch:1c | Note:Unlabeled AAs were used for total AA (FigS2.D) RAW_FILE_NAME=bms-fu-2.cdf SUBJECT_SAMPLE_FACTORS - BM_supernatant_5FU_3 Source_Name[gating]:Bone marrow supernatant | Treatment:5 FU 2 weeks | Batch:1c | Note:Unlabeled AAs were used for total AA (FigS2.D) RAW_FILE_NAME=bms-fu-3.cdf SUBJECT_SAMPLE_FACTORS - BM_supernatant_Ctrl_1 Source_Name[gating]:Bone marrow supernatant | Treatment:HSC Basal | Batch:1c | Note:Unlabeled AAs were used for total AA (FigS2.D) RAW_FILE_NAME=bms-ct-1.cdf SUBJECT_SAMPLE_FACTORS - BM_supernatant_Ctrl_2 Source_Name[gating]:Bone marrow supernatant | Treatment:HSC Basal | Batch:1c | Note:Unlabeled AAs were used for total AA (FigS2.D) RAW_FILE_NAME=bms-ct-2.cdf SUBJECT_SAMPLE_FACTORS - BM_supernatant_Ctrl_3 Source_Name[gating]:Bone marrow supernatant | Treatment:HSC Basal | Batch:1c | Note:Unlabeled AAs were used for total AA (FigS2.D) RAW_FILE_NAME=bms-ct-3.cdf SUBJECT_SAMPLE_FACTORS - HSC_5FU_1 Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:5 FU 2 weeks, [13C, 15N] AAs were incorporated for 1 h | Batch:1c | Note:[13C, 15N] AAs were used for AA uptake (Fig2.B, Fig5M); unlabeled AAs and TCA substrate were used for total AA levels (Fig2.C, Fig5K) and TCA substrate levels (Fig2.K, Fig5J). RAW_FILE_NAME=hsc-fu-1.cdf SUBJECT_SAMPLE_FACTORS - HSC_5FU_2 Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:5 FU 2 weeks, [13C, 15N] AAs were incorporated for 1 h | Batch:1c | Note:[13C, 15N] AAs were used for AA uptake (Fig2.B, Fig5M); unlabeled AAs and TCA substrate were used for total AA levels (Fig2.C, Fig5K) and TCA substrate levels (Fig2.K, Fig5J). RAW_FILE_NAME=hsc-fu-2.cdf SUBJECT_SAMPLE_FACTORS - HSC_5FU_3 Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:5 FU 2 weeks, [13C, 15N] AAs were incorporated for 1 h | Batch:1c | Note:[13C, 15N] AAs were used for AA uptake (Fig2.B, Fig5M); unlabeled AAs and TCA substrate were used for total AA levels (Fig2.C, Fig5K) and TCA substrate levels (Fig2.K, Fig5J). RAW_FILE_NAME=hsc-fu-3.cdf SUBJECT_SAMPLE_FACTORS - HSC_5FU_0h_1 Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:5 FU 2 weeks, [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 (Fig2.D). RAW_FILE_NAME=hsc-fu-0h-1.cdf SUBJECT_SAMPLE_FACTORS - HSC_5FU_0h_2 Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:5 FU 2 weeks, [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 (Fig2.D). RAW_FILE_NAME=hsc-fu-0h-2.cdf SUBJECT_SAMPLE_FACTORS - HSC_5FU_0h_3 Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:5 FU 2 weeks, [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 (Fig2.D). RAW_FILE_NAME=hsc-fu-0h-3.cdf SUBJECT_SAMPLE_FACTORS - HSC_5FU_12h_1 Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:5 FU 2 weeks, [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 (Fig2.D). RAW_FILE_NAME=hsc-fu-12h-1.cdf SUBJECT_SAMPLE_FACTORS - HSC_5FU_12h_2 Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:5 FU 2 weeks, [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 (Fig2.D). RAW_FILE_NAME=hsc-fu-12h-2.cdf SUBJECT_SAMPLE_FACTORS - HSC_5FU_12h_3 Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:5 FU 2 weeks, [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 (Fig2.D). RAW_FILE_NAME=hsc-fu-12h-3.cdf SUBJECT_SAMPLE_FACTORS - HSC_5FU_6h_1 Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:5 FU 2 weeks, [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 (Fig2.D). RAW_FILE_NAME=hsc-fu-6h-1.cdf SUBJECT_SAMPLE_FACTORS - HSC_5FU_6h_2 Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:5 FU 2 weeks, [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 (Fig2.D). RAW_FILE_NAME=hsc-fu-6h-2.cdf SUBJECT_SAMPLE_FACTORS - HSC_5FU_6h_3 Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:5 FU 2 weeks, [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 (Fig2.D). RAW_FILE_NAME=hsc-fu-6h-3.cdf SUBJECT_SAMPLE_FACTORS - HSC_Cultured_1 Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:Ex vivo culture 2 weeks, [13C, 15N] AAs were incorporated for 1 h | Batch:1c | Note:[13C, 15N] AAs were used for AA uptake (Fig2.B); unlabeled AAs and TCA substrate were used for total AA levels (Fig2.C) and TCA substrate levels (Fig2.K). RAW_FILE_NAME=hsc-cultured-1.cdf SUBJECT_SAMPLE_FACTORS - HSC_Cultured_2 Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:Ex vivo culture 2 weeks, [13C, 15N] AAs were incorporated for 1 h | Batch:1c | Note:[13C, 15N] AAs were used for AA uptake (Fig2.B); unlabeled AAs and TCA substrate were used for total AA levels (Fig2.C) and TCA substrate levels (Fig2.K). RAW_FILE_NAME=hsc-cultured-2.cdf SUBJECT_SAMPLE_FACTORS - HSC_Cultured_3 Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:Ex vivo culture 2 weeks, [13C, 15N] AAs were incorporated for 1 h | Batch:1c | Note:[13C, 15N] AAs were used for AA uptake (Fig2.B); unlabeled AAs and TCA substrate were used for total AA levels (Fig2.C) and TCA substrate levels (Fig2.K). RAW_FILE_NAME=hsc-cultured-3.cdf SUBJECT_SAMPLE_FACTORS - HSC_Cultured_0h_1 Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:Ex vivo culture 2 weeks, [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 (Fig2.D). RAW_FILE_NAME=hsc-cultured-0h-1.cdf SUBJECT_SAMPLE_FACTORS - HSC_Cultured_0h_2 Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:Ex vivo culture 2 weeks, [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 (Fig2.D). RAW_FILE_NAME=hsc-cultured-0h-2.cdf SUBJECT_SAMPLE_FACTORS - HSC_Cultured_0h_3 Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:Ex vivo culture 2 weeks, [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 (Fig2.D). RAW_FILE_NAME=hsc-cultured-0h-3.cdf SUBJECT_SAMPLE_FACTORS - HSC_Cultured_6h_1 Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:Ex vivo culture 2 weeks, [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 (Fig2.D). RAW_FILE_NAME=hsc-cultured-6h-1.cdf SUBJECT_SAMPLE_FACTORS - HSC_Cultured_12h_1 Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:Ex vivo culture 2 weeks, [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 (Fig2.D). RAW_FILE_NAME=hsc-cultured-12h-1.cdf SUBJECT_SAMPLE_FACTORS - HSC_Cultured_12h_2 Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:Ex vivo culture 2 weeks, [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 (Fig2.D). RAW_FILE_NAME=hsc-cultured-12h-2.cdf SUBJECT_SAMPLE_FACTORS - HSC_Cultured_12h_3 Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:Ex vivo culture 2 weeks, [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 (Fig2.D). RAW_FILE_NAME=hsc-cultured-12h-3.cdf SUBJECT_SAMPLE_FACTORS - HSC_Cultured_6h_2 Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:Ex vivo culture 2 weeks, [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 (Fig2.D). RAW_FILE_NAME=hsc-cultured-6h-2.cdf SUBJECT_SAMPLE_FACTORS - HSC_Cultured_6h_3 Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:Ex vivo culture 2 weeks, [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 (Fig2.D). RAW_FILE_NAME=hsc-cultured-6h-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 - HSC_Ctrl_1 Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:HSC Basal or Ctrl, [13C, 15N] AAs were incorporated for 1 h | Batch:1c | Note:[13C, 15N] AAs were used for AA uptake (Fig2.B, Fig5M); unlabeled AAs and TCA substrate were used for total AA levels (Fig2.C, Fig5K) and TCA substrate levels (Fig2.K, Fig5J). RAW_FILE_NAME=hsc-ct-1.cdf SUBJECT_SAMPLE_FACTORS - HSC_Ctrl_2 Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:HSC Basal or Ctrl, [13C, 15N] AAs were incorporated for 1 h | Batch:1c | Note:[13C, 15N] AAs were used for AA uptake (Fig2.B, Fig5M); unlabeled AAs and TCA substrate were used for total AA levels (Fig2.C, Fig5K) and TCA substrate levels (Fig2.K, Fig5J). RAW_FILE_NAME=hsc-ct-2.cdf SUBJECT_SAMPLE_FACTORS - HSC_Ctrl_3 Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:HSC Basal or Ctrl, [13C, 15N] AAs were incorporated for 1 h | Batch:1c | Note:[13C, 15N] AAs were used for AA uptake (Fig2.B, Fig5M); unlabeled AAs and TCA substrate were used for total AA levels (Fig2.C, Fig5K) and TCA substrate levels (Fig2.K, Fig5J). RAW_FILE_NAME=hsc-ct-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 BM_supernatant_5FU_1 BM_supernatant_5FU_2 BM_supernatant_5FU_3 BM_supernatant_Ctrl_1 BM_supernatant_Ctrl_2 BM_supernatant_Ctrl_3 HSC_5FU_1 HSC_5FU_2 HSC_5FU_3 HSC_5FU_0h_1 HSC_5FU_0h_2 HSC_5FU_0h_3 HSC_5FU_12h_1 HSC_5FU_12h_2 HSC_5FU_12h_3 HSC_5FU_6h_1 HSC_5FU_6h_2 HSC_5FU_6h_3 HSC_Cultured_1 HSC_Cultured_2 HSC_Cultured_3 HSC_Cultured_0h_1 HSC_Cultured_0h_2 HSC_Cultured_0h_3 HSC_Cultured_6h_1 HSC_Cultured_12h_1 HSC_Cultured_12h_2 HSC_Cultured_12h_3 HSC_Cultured_6h_2 HSC_Cultured_6h_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 HSC_Ctrl_1 HSC_Ctrl_2 HSC_Ctrl_3 Factors Source_Name[gating]:Bone marrow supernatant | Treatment:5 FU 2 weeks | Batch:1c | Note:Unlabeled AAs were used for total AA (FigS2.D) Source_Name[gating]:Bone marrow supernatant | Treatment:5 FU 2 weeks | Batch:1c | Note:Unlabeled AAs were used for total AA (FigS2.D) Source_Name[gating]:Bone marrow supernatant | Treatment:5 FU 2 weeks | Batch:1c | Note:Unlabeled AAs were used for total AA (FigS2.D) Source_Name[gating]:Bone marrow supernatant | Treatment:HSC Basal | Batch:1c | Note:Unlabeled AAs were used for total AA (FigS2.D) Source_Name[gating]:Bone marrow supernatant | Treatment:HSC Basal | Batch:1c | Note:Unlabeled AAs were used for total AA (FigS2.D) Source_Name[gating]:Bone marrow supernatant | Treatment:HSC Basal | Batch:1c | Note:Unlabeled AAs were used for total AA (FigS2.D) Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:5 FU 2 weeks, [13C, 15N] AAs were incorporated for 1 h | Batch:1c | Note:[13C, 15N] AAs were used for AA uptake (Fig2.B, Fig5M); unlabeled AAs and TCA substrate were used for total AA levels (Fig2.C, Fig5K) and TCA substrate levels (Fig2.K, Fig5J). Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:5 FU 2 weeks, [13C, 15N] AAs were incorporated for 1 h | Batch:1c | Note:[13C, 15N] AAs were used for AA uptake (Fig2.B, Fig5M); unlabeled AAs and TCA substrate were used for total AA levels (Fig2.C, Fig5K) and TCA substrate levels (Fig2.K, Fig5J). Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:5 FU 2 weeks, [13C, 15N] AAs were incorporated for 1 h | Batch:1c | Note:[13C, 15N] AAs were used for AA uptake (Fig2.B, Fig5M); unlabeled AAs and TCA substrate were used for total AA levels (Fig2.C, Fig5K) and TCA substrate levels (Fig2.K, Fig5J). Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:5 FU 2 weeks, [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 (Fig2.D). Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:5 FU 2 weeks, [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 (Fig2.D). Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:5 FU 2 weeks, [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 (Fig2.D). Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:5 FU 2 weeks, [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 (Fig2.D). Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:5 FU 2 weeks, [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 (Fig2.D). Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:5 FU 2 weeks, [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 (Fig2.D). Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:5 FU 2 weeks, [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 (Fig2.D). Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:5 FU 2 weeks, [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 (Fig2.D). Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:5 FU 2 weeks, [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 (Fig2.D). Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:Ex vivo culture 2 weeks, [13C, 15N] AAs were incorporated for 1 h | Batch:1c | Note:[13C, 15N] AAs were used for AA uptake (Fig2.B); unlabeled AAs and TCA substrate were used for total AA levels (Fig2.C) and TCA substrate levels (Fig2.K). Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:Ex vivo culture 2 weeks, [13C, 15N] AAs were incorporated for 1 h | Batch:1c | Note:[13C, 15N] AAs were used for AA uptake (Fig2.B); unlabeled AAs and TCA substrate were used for total AA levels (Fig2.C) and TCA substrate levels (Fig2.K). Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:Ex vivo culture 2 weeks, [13C, 15N] AAs were incorporated for 1 h | Batch:1c | Note:[13C, 15N] AAs were used for AA uptake (Fig2.B); unlabeled AAs and TCA substrate were used for total AA levels (Fig2.C) and TCA substrate levels (Fig2.K). Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:Ex vivo culture 2 weeks, [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 (Fig2.D). Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:Ex vivo culture 2 weeks, [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 (Fig2.D). Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:Ex vivo culture 2 weeks, [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 (Fig2.D). Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:Ex vivo culture 2 weeks, [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 (Fig2.D). Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:Ex vivo culture 2 weeks, [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 (Fig2.D). Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:Ex vivo culture 2 weeks, [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 (Fig2.D). Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:Ex vivo culture 2 weeks, [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 (Fig2.D). Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:Ex vivo culture 2 weeks, [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 (Fig2.D). Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:Ex vivo culture 2 weeks, [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 (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 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+ CD48– CD150+ (HSC) | Treatment:HSC Basal or Ctrl, [13C, 15N] AAs were incorporated for 1 h | Batch:1c | Note:[13C, 15N] AAs were used for AA uptake (Fig2.B, Fig5M); unlabeled AAs and TCA substrate were used for total AA levels (Fig2.C, Fig5K) and TCA substrate levels (Fig2.K, Fig5J). Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:HSC Basal or Ctrl, [13C, 15N] AAs were incorporated for 1 h | Batch:1c | Note:[13C, 15N] AAs were used for AA uptake (Fig2.B, Fig5M); unlabeled AAs and TCA substrate were used for total AA levels (Fig2.C, Fig5K) and TCA substrate levels (Fig2.K, Fig5J). Source_Name[gating]:Lin– Scal1+ c-Kit+ CD48– CD150+ (HSC) | Treatment:HSC Basal or Ctrl, [13C, 15N] AAs were incorporated for 1 h | Batch:1c | Note:[13C, 15N] AAs were used for AA uptake (Fig2.B, Fig5M); unlabeled AAs and TCA substrate were used for total AA levels (Fig2.C, Fig5K) and TCA substrate levels (Fig2.K, Fig5J). [13C_ 15N]Alanine_ 2TBDMS 66700.0000 68400.0000 68400.0000 6670000.0000 6840000.0000 6840000.0000 6110000.0000 6030000.0000 6350000.0000 5750000.0000 5440000.0000 5620000.0000 69100.0000 66300.0000 67100.0000 6910000.0000 6630000.0000 6710000.0000 5410000.0000 6170000.0000 6280000.0000 6040000.0000 5620000.0000 5530000.0000 6500000.0000 7200000.0000 6290000.0000 4760000.0000 5250000.0000 4820000.0000 5550000.0000 6110000.0000 5630000.0000 61300.0000 60400.0000 63200.0000 [13C_ 15N]Aspartic acid_ 3TBDMS 96300.0000 113000.0000 128000.0000 9630000.0000 11300000.0000 12800000.0000 4560000.0000 5120000.0000 4120000.0000 6430000.0000 6120000.0000 6020000.0000 102000.0000 118000.0000 124000.0000 10200000.0000 11800000.0000 12400000.0000 5640000.0000 4850000.0000 4120000.0000 4320000.0000 5120000.0000 5580000.0000 8690000.0000 9120000.0000 8900000.0000 2590000.0000 4080000.0000 3610000.0000 4330000.0000 6320000.0000 5630000.0000 78400.0000 86300.0000 71300.0000 [13C_ 15N]Glutamic acid_ 3TBDMS 74200.0000 71200.0000 73100.0000 7420000.0000 7120000.0000 7310000.0000 2130000.0000 2440000.0000 2320000.0000 4220000.0000 3860000.0000 3560000.0000 68600.0000 73200.0000 66400.0000 6860000.0000 7320000.0000 6640000.0000 3300000.0000 1860000.0000 1520000.0000 1750000.0000 3100000.0000 3290000.0000 3090000.0000 2590000.0000 2620000.0000 440000.0000 657000.0000 588000.0000 1750000.0000 1500000.0000 1650000.0000 32500.0000 31200.0000 35200.0000 [13C_ 15N]Glycine_ 2TBDMS 87200.0000 68500.0000 79900.0000 7030000.0000 6780000.0000 6500000.0000 4580000.0000 3840000.0000 4510000.0000 4700000.0000 5880000.0000 5200000.0000 88600.0000 71100.0000 70800.0000 6960000.0000 6310000.0000 7670000.0000 5470000.0000 3390000.0000 5930000.0000 4640000.0000 5520000.0000 4410000.0000 6490000.0000 5780000.0000 4740000.0000 3690000.0000 2190000.0000 2590000.0000 3180000.0000 5300000.0000 3670000.0000 53400.0000 53500.0000 66900.0000 [13C_ 15N]Histidine_ 3TBDMS 54100.0000 48700.0000 45200.0000 5410000.0000 4870000.0000 4520000.0000 3860000.0000 3530000.0000 3430000.0000 4220000.0000 4870000.0000 3350000.0000 51000.0000 45200.0000 47600.0000 5100000.0000 4520000.0000 4760000.0000 4260000.0000 3120000.0000 3350000.0000 3420000.0000 3660000.0000 4200000.0000 3060000.0000 3480000.0000 3500000.0000 1120000.0000 936000.0000 853000.0000 2150000.0000 2230000.0000 2530000.0000 42300.0000 39900.0000 38400.0000 [13C_ 15N]Isoleucine_ 2TBDMS 183000.0000 163000.0000 163000.0000 18300000.0000 16300000.0000 16300000.0000 10700000.0000 10400000.0000 9950000.0000 13500000.0000 15200000.0000 14700000.0000 174000.0000 163000.0000 152000.0000 17400000.0000 16300000.0000 15200000.0000 15200000.0000 12300000.0000 12100000.0000 13200000.0000 14300000.0000 14400000.0000 9450000.0000 13200000.0000 11500000.0000 3430000.0000 3140000.0000 4050000.0000 6520000.0000 6120000.0000 7130000.0000 146000.0000 122000.0000 130000.0000 [13C_ 15N]Leucine_ 2TBDMS 183000.0000 213000.0000 203000.0000 18300000.0000 21300000.0000 20300000.0000 8560000.0000 9530000.0000 7820000.0000 15200000.0000 13300000.0000 13200000.0000 232000.0000 210000.0000 224000.0000 23200000.0000 21000000.0000 22400000.0000 12000000.0000 8100000.0000 7430000.0000 8030000.0000 14200000.0000 13300000.0000 10300000.0000 14900000.0000 12900000.0000 4660000.0000 4060000.0000 4370000.0000 8610000.0000 8120000.0000 8310000.0000 133000.0000 154000.0000 117000.0000 [13C_ 15N]Lysine_ 3TBDMS 74600.0000 84600.0000 78400.0000 7460000.0000 8460000.0000 7840000.0000 4230000.0000 3550000.0000 3880000.0000 5220000.0000 4880000.0000 6520000.0000 84200.0000 85200.0000 81200.0000 8420000.0000 8520000.0000 8120000.0000 5560000.0000 4110000.0000 3880000.0000 3600000.0000 4760000.0000 5130000.0000 5630000.0000 7840000.0000 6220000.0000 2980000.0000 3580000.0000 3040000.0000 5650000.0000 4520000.0000 5970000.0000 58400.0000 51200.0000 56300.0000 [13C_ 15N]Methionine_ 2TBDMS 183000.0000 173000.0000 193000.0000 18300000.0000 17300000.0000 19300000.0000 12100000.0000 11700000.0000 13000000.0000 14300000.0000 15300000.0000 14400000.0000 163000.0000 150000.0000 168000.0000 16300000.0000 15000000.0000 16800000.0000 14400000.0000 12000000.0000 11000000.0000 10300000.0000 12300000.0000 12900000.0000 9660000.0000 12800000.0000 15500000.0000 7580000.0000 6760000.0000 7070000.0000 8510000.0000 8630000.0000 8510000.0000 152000.0000 130000.0000 142000.0000 [13C_ 15N]Phenylalanine_ 2TBDMS 132000.0000 142000.0000 125000.0000 13200000.0000 14200000.0000 12500000.0000 6350000.0000 6120000.0000 6240000.0000 11000000.0000 8960000.0000 9120000.0000 124000.0000 142000.0000 142000.0000 12400000.0000 14200000.0000 14200000.0000 9650000.0000 7450000.0000 6320000.0000 7120000.0000 11000000.0000 10600000.0000 11400000.0000 15900000.0000 11200000.0000 5630000.0000 4620000.0000 4530000.0000 8530000.0000 8420000.0000 7510000.0000 115000.0000 105000.0000 125000.0000 [13C_ 15N]Proline_ 2TBDMS 143000.0000 132000.0000 133000.0000 14300000.0000 13200000.0000 13300000.0000 8630000.0000 7520000.0000 8880000.0000 11300000.0000 10400000.0000 11000000.0000 152000.0000 163000.0000 152000.0000 15200000.0000 16300000.0000 15200000.0000 13200000.0000 8540000.0000 7430000.0000 7040000.0000 11500000.0000 12500000.0000 7570000.0000 8770000.0000 7560000.0000 2800000.0000 1980000.0000 2880000.0000 6120000.0000 5250000.0000 5150000.0000 78500.0000 86300.0000 88500.0000 [13C_ 15N]Serine_ 3TBDMS 132000.0000 125000.0000 130000.0000 13200000.0000 12500000.0000 13000000.0000 7540000.0000 8510000.0000 8000000.0000 10200000.0000 9860000.0000 10400000.0000 142000.0000 140000.0000 133000.0000 14200000.0000 14000000.0000 13300000.0000 11500000.0000 7540000.0000 8000000.0000 7850000.0000 10600000.0000 11100000.0000 9600000.0000 8340000.0000 9860000.0000 6160000.0000 6690000.0000 6980000.0000 7330000.0000 7230000.0000 8890000.0000 84200.0000 75100.0000 80100.0000 [13C_ 15N]Threonine_ 3TBDMS 17500.0000 18700.0000 20000.0000 1750000.0000 1870000.0000 2000000.0000 1030000.0000 1190000.0000 1030000.0000 1520000.0000 1520000.0000 1440000.0000 18800.0000 19500.0000 19500.0000 1880000.0000 1950000.0000 1950000.0000 1550000.0000 1150000.0000 1200000.0000 1110000.0000 1760000.0000 1450000.0000 1660000.0000 1580000.0000 2080000.0000 795000.0000 889000.0000 885000.0000 1130000.0000 1110000.0000 1250000.0000 12500.0000 14300.0000 14400.0000 [13C_ 15N]Tyrosine_ 3TBDMS 855000.0000 812000.0000 832000.0000 85500000.0000 81200000.0000 83200000.0000 41200000.0000 39900000.0000 37400000.0000 63300000.0000 61000000.0000 61000000.0000 874000.0000 833000.0000 842000.0000 87400000.0000 83300000.0000 84200000.0000 58400000.0000 32600000.0000 30100000.0000 33500000.0000 60200000.0000 61600000.0000 47400000.0000 54000000.0000 44500000.0000 16900000.0000 23400000.0000 25200000.0000 35400000.0000 32200000.0000 30400000.0000 523000.0000 488000.0000 512000.0000 [13C_ 15N]Valine_ 2TBDMS 63200.0000 61300.0000 64300.0000 63200000.0000 61300000.0000 64300000.0000 45100000.0000 41200000.0000 43200000.0000 53600000.0000 55100000.0000 54100000.0000 71300.0000 73000.0000 68200.0000 71300000.0000 73000000.0000 68200000.0000 43200000.0000 36300000.0000 41100000.0000 40100000.0000 53400000.0000 51200000.0000 51600000.0000 41200000.0000 45500000.0000 15500000.0000 17200000.0000 14000000.0000 28300000.0000 31400000.0000 25200000.0000 41200.0000 43200.0000 41300.0000 [13C]Alpha_Ketoglutarate 6380000.0000 6250000.0000 6110000.0000 18400.0000 17600.0000 18400.0000 18800.0000 22000.0000 21400.0000 19900.0000 21000.0000 20500.0000 6130000.0000 6250000.0000 6260000.0000 15200.0000 14200.0000 15200.0000 16600.0000 18600.0000 18400.0000 18000.0000 17400.0000 17000.0000 25500.0000 30200.0000 27200.0000 83400.0000 64600.0000 82700.0000 41200.0000 41200.0000 49700.0000 4010000.0000 4120000.0000 4180000.0000 [13C]Citrate 14400000.0000 14200000.0000 15200000.0000 32900.0000 30500.0000 31800.0000 48900.0000 42200.0000 45900.0000 40200.0000 38600.0000 34200.0000 16300000.0000 16200000.0000 16200000.0000 31000.0000 35200.0000 32800.0000 36600.0000 41100.0000 40300.0000 42100.0000 37100.0000 35400.0000 50500.0000 60800.0000 62000.0000 192000.0000 194000.0000 212000.0000 115000.0000 108000.0000 125000.0000 8530000.0000 7950000.0000 7680000.0000 [13C]Fumatate 62200000.0000 62300000.0000 62300000.0000 123000.0000 112000.0000 132000.0000 149000.0000 144000.0000 142000.0000 142000.0000 132000.0000 113000.0000 67100000.0000 65600000.0000 62700000.0000 142000.0000 152000.0000 136000.0000 132000.0000 152000.0000 163000.0000 156000.0000 152000.0000 163000.0000 209000.0000 181000.0000 226000.0000 290000.0000 313000.0000 313000.0000 256000.0000 243000.0000 264000.0000 41100000.0000 40100000.0000 42000000.0000 [13C]Malate 21600000.0000 24000000.0000 22900000.0000 65300.0000 60300.0000 70300.0000 78600.0000 71200.0000 78600.0000 74200.0000 72300.0000 74000.0000 22000000.0000 24900000.0000 23900000.0000 70300.0000 67600.0000 61200.0000 73200.0000 76600.0000 77600.0000 74100.0000 72300.0000 70200.0000 96700.0000 86700.0000 76100.0000 114000.0000 119000.0000 131000.0000 92700.0000 83300.0000 105000.0000 11700000.0000 9260000.0000 10400000.0000 [13C]Succinate 67300000.0000 61300000.0000 65300000.0000 112000.0000 102000.0000 98600.0000 132000.0000 142000.0000 120000.0000 120000.0000 112000.0000 128000.0000 63500000.0000 64600000.0000 65600000.0000 102000.0000 89700.0000 110000.0000 112000.0000 141000.0000 137000.0000 140000.0000 120000.0000 125000.0000 142000.0000 183000.0000 153000.0000 293000.0000 267000.0000 244000.0000 215000.0000 204000.0000 215000.0000 41700000.0000 40900000.0000 42900000.0000 Alanine_ 2TBDMS 317000000.0000 322000000.0000 316000000.0000 83800000.0000 82100000.0000 68100000.0000 5200000.0000 4720000.0000 5210000.0000 6220000.0000 5860000.0000 4120000.0000 1230000.0000 1590000.0000 1420000.0000 Alpha_Ketoglutarate 6380000.0000 6250000.0000 6110000.0000 6130000.0000 6250000.0000 6260000.0000 4010000.0000 4120000.0000 4180000.0000 Aspartic acid_ 3TBDMS 3190000000.0000 3220000000.0000 3150000000.0000 858000000.0000 922000000.0000 733000000.0000 154000000.0000 135000000.0000 137000000.0000 129000000.0000 152000000.0000 149000000.0000 42100000.0000 45200000.0000 47600000.0000 Citrate 14400000.0000 14200000.0000 15200000.0000 16300000.0000 16200000.0000 16200000.0000 8530000.0000 7950000.0000 7680000.0000 Fumatate 62200000.0000 62300000.0000 62300000.0000 67100000.0000 65600000.0000 62700000.0000 41100000.0000 40100000.0000 42000000.0000 Glutamic acid_ 3TBDMS 3800000000.0000 3910000000.0000 3630000000.0000 1770000000.0000 1870000000.0000 1590000000.0000 132000000.0000 142000000.0000 174000000.0000 159000000.0000 170000000.0000 188000000.0000 42100000.0000 40300000.0000 43600000.0000 Glutamine_ 3TBDMS 325000000.0000 357000000.0000 415000000.0000 158000000.0000 135000000.0000 162000000.0000 1740000.0000 1860000.0000 1520000.0000 1740000.0000 1400000.0000 1590000.0000 386000.0000 412000.0000 432000.0000 Glycine_ 2TBDMS 14900000000.0000 13400000000.0000 13300000000.0000 3280000000.0000 3250000000.0000 2740000000.0000 542000000.0000 612000000.0000 586000000.0000 486000000.0000 474000000.0000 452000000.0000 253000000.0000 214000000.0000 253000000.0000 Histidine_ 3TBDMS 221000000.0000 228000000.0000 216000000.0000 78400000.0000 89300000.0000 62500000.0000 284000.0000 320000.0000 218000.0000 205000.0000 353000.0000 188000.0000 52100.0000 41900.0000 52900.0000 Hydroxyproline_ (E)-_ 3TBDMS 94300000.0000 99400000.0000 99000000.0000 45500000.0000 51300000.0000 23200000.0000 4760000.0000 5200000.0000 5340000.0000 4890000.0000 5120000.0000 5010000.0000 1190000.0000 1390000.0000 1240000.0000 Isoleucine_ 2TBDMS 2120000000.0000 2140000000.0000 2100000000.0000 429000000.0000 466000000.0000 405000000.0000 15400000.0000 17400000.0000 18400000.0000 15600000.0000 17400000.0000 17400000.0000 5410000.0000 5120000.0000 5340000.0000 L-Cysteine_ 3TBDMS 16700000.0000 12300000.0000 14500000.0000 2960000.0000 3020000.0000 2670000.0000 3250000.0000 2840000.0000 3100000.0000 2860000.0000 3700000.0000 3190000.0000 686000.0000 854000.0000 774000.0000 Leucine_ 2TBDMS 1080000000.0000 1170000000.0000 1020000000.0000 666000000.0000 459000000.0000 710000000.0000 41900000.0000 45800000.0000 43900000.0000 39800000.0000 41900000.0000 43400000.0000 12000000.0000 13900000.0000 14100000.0000 Lysine_ 3TBDMS 2140000000.0000 2180000000.0000 2110000000.0000 470000000.0000 529000000.0000 418000000.0000 15200000.0000 18600000.0000 14400000.0000 15200000.0000 12000000.0000 13400000.0000 4220000.0000 3420000.0000 4180000.0000 Malate 21600000.0000 24000000.0000 22900000.0000 22000000.0000 24900000.0000 23900000.0000 11700000.0000 9260000.0000 10400000.0000 Methionine_ 2TBDMS 1270000000.0000 1170000000.0000 1150000000.0000 293000000.0000 319000000.0000 278000000.0000 12300000.0000 14200000.0000 13400000.0000 15200000.0000 14700000.0000 13500000.0000 3520000.0000 3100000.0000 3370000.0000 Phenylalanine_ 2TBDMS 2110000000.0000 2130000000.0000 2590000000.0000 420000000.0000 458000000.0000 395000000.0000 6520000.0000 4320000.0000 6320000.0000 7410000.0000 6530000.0000 6850000.0000 1860000.0000 2100000.0000 2100000.0000 Proline_ 2TBDMS 1610000000.0000 1410000000.0000 1590000000.0000 371000000.0000 464000000.0000 421000000.0000 65500000.0000 58200000.0000 87000000.0000 87500000.0000 63200000.0000 74300000.0000 10200000.0000 13200000.0000 12500000.0000 Serine_ 3TBDMS 3260000000.0000 3300000000.0000 3210000000.0000 836000000.0000 930000000.0000 753000000.0000 52200000.0000 51700000.0000 56000000.0000 61500000.0000 58200000.0000 60200000.0000 18600000.0000 21800000.0000 20900000.0000 Succinate 67300000.0000 61300000.0000 65300000.0000 63500000.0000 64600000.0000 65600000.0000 41700000.0000 40900000.0000 42900000.0000 Threonine_ 3TBDMS 329000000.0000 335000000.0000 320000000.0000 90100000.0000 131000000.0000 70100000.0000 8540000.0000 7420000.0000 6230000.0000 4240000.0000 7550000.0000 8630000.0000 2860000.0000 2510000.0000 2150000.0000 Tyrosine_ 3TBDMS 5310000000.0000 4360000000.0000 4240000000.0000 1580000000.0000 1620000000.0000 1500000000.0000 18400000.0000 21600000.0000 21000000.0000 23600000.0000 24700000.0000 22000000.0000 4720000.0000 5820000.0000 6340000.0000 Valine_ 2TBDMS 3330000000.0000 3380000000.0000 3260000000.0000 883000000.0000 947000000.0000 841000000.0000 52100000.0000 42300000.0000 51200000.0000 41100000.0000 51200000.0000 48600000.0000 11200000.0000 13200000.0000 14200000.0000 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