#METABOLOMICS WORKBENCH kevin.cho@wustl.edu_20241126_081352 DATATRACK_ID:5407 STUDY_ID:ST003598 ANALYSIS_ID:AN005911 PROJECT_ID:PR002226 VERSION 1 CREATED_ON November 27, 2024, 7:47 am #PROJECT PR:PROJECT_TITLE Nuclear Factor I A (NFIA) regulates articular chondrocyte fatty acid metabolism PR:PROJECT_TITLE and joint homeostasis PR:PROJECT_SUMMARY Osteoarthritis (OA) has recently been recognized as a joint disease with an PR:PROJECT_SUMMARY etiology partially rooted in metabolic dysfunction, yet the underlying PR:PROJECT_SUMMARY mechanism(s) in this context are not determined, limiting opportunities to PR:PROJECT_SUMMARY develop therapeutic treatments. In this study, we utilized a multi-omics PR:PROJECT_SUMMARY approach combining RNA-seq, ATAC-seq, MRE-seq, and metabolomics to reveal that PR:PROJECT_SUMMARY OA articular chondrocytes induced by imbalanced Transforming Growth Factor β PR:PROJECT_SUMMARY 1/Bone Morphogenetic Protein 2 (TGF-β/BMP) signaling, possess increased fatty PR:PROJECT_SUMMARY acid synthesis and oxidation processes regulated by Nuclear Factor I A (NFIA) PR:PROJECT_SUMMARY upregulation PR:INSTITUTE Washington University in St. Louis PR:DEPARTMENT Orthopaedic Surgery, Chemistry, Medicine PR:LABORATORY Shen Laboratory and Patti Laboratory PR:LAST_NAME Cho PR:FIRST_NAME Kevin PR:ADDRESS 1 Brookings Drive PR:EMAIL kevin.cho@wustl.edu PR:PHONE 314-935-8813 #STUDY ST:STUDY_TITLE Nuclear Factor I A (NFIA) Regulates Articular Chondrocyte Fatty Acid Metabolism ST:STUDY_TITLE and Joint Homeostasis ST:STUDY_SUMMARY Osteoarthritis (OA) has recently been recognized as a joint disease with an ST:STUDY_SUMMARY etiology partially rooted in metabolic dysfunction, yet the underlying ST:STUDY_SUMMARY mechanism(s) in this context are not determined, limiting opportunities to ST:STUDY_SUMMARY develop therapeutic treatments. In this study, we utilized a multi-omics ST:STUDY_SUMMARY approach combining RNA-seq, ATAC-seq, MRE-seq, and metabolomics to reveal that ST:STUDY_SUMMARY OA articular chondrocytes induced by imbalanced Transforming Growth Factor β ST:STUDY_SUMMARY 1/Bone Morphogenetic Protein 2 (TGF-β/BMP) signaling, possess increased fatty ST:STUDY_SUMMARY acid synthesis and oxidation processes regulated by Nuclear Factor I A (NFIA) ST:STUDY_SUMMARY upregulation ST:INSTITUTE Washington University in St. Louis ST:DEPARTMENT Orthopaedic Surgery, Chemistry, Medicine ST:LABORATORY Shen Laboratory and Patti Laboratory ST:LAST_NAME Cho ST:FIRST_NAME Kevin ST:ADDRESS 1 Brookings Drive ST:EMAIL kevin.cho@wustl.edu ST:PHONE 314-935-8813 #SUBJECT SU:SUBJECT_TYPE Mammal SU:SUBJECT_SPECIES Mus musculus; Homo sapiens SU:TAXONOMY_ID 10090; 9606 #FACTORS #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Raw file names and additional sample data SUBJECT_SAMPLE_FACTORS M-Chon-01 M_TGFb_Gln_1 Sample source:Mice Chondrocytes | Genotype:WT | Treatment:TGFb1 Label=U-13C-Glutamine; RAW_FILE_NAME(Raw file name)=M-Chon-01.mzML SUBJECT_SAMPLE_FACTORS M-Chon-02 M_TGFb_Gln_2 Sample source:Mice Chondrocytes | Genotype:WT | Treatment:TGFb1 Label=U-13C-Glutamine; RAW_FILE_NAME(Raw file name)=M-Chon-02.mzML SUBJECT_SAMPLE_FACTORS M-Chon-03 M_TGFb_Gln_3 Sample source:Mice Chondrocytes | Genotype:WT | Treatment:TGFb1 Label=U-13C-Glutamine; RAW_FILE_NAME(Raw file name)=M-Chon-03.mzML SUBJECT_SAMPLE_FACTORS M-Chon-04 M_BMP_Gln_1 Sample source:Mice Chondrocytes | Genotype:WT | Treatment:BMP2 Label=U-13C-Glutamine; RAW_FILE_NAME(Raw file name)=M-Chon-04.mzML SUBJECT_SAMPLE_FACTORS M-Chon-05 M_BMP_Gln_2 Sample source:Mice Chondrocytes | Genotype:WT | Treatment:BMP2 Label=U-13C-Glutamine; RAW_FILE_NAME(Raw file name)=M-Chon-05.mzML SUBJECT_SAMPLE_FACTORS M-Chon-06 M_BMP_Gln_3 Sample source:Mice Chondrocytes | Genotype:WT | Treatment:BMP2 Label=U-13C-Glutamine; RAW_FILE_NAME(Raw file name)=M-Chon-06.mzML SUBJECT_SAMPLE_FACTORS M-Chon-07 M_TGFb_PA_1 Sample source:Mice Chondrocytes | Genotype:WT | Treatment:TGFb1 Label=U-13C-Palmitate; RAW_FILE_NAME(Raw file name)=M-Chon-07.mzML SUBJECT_SAMPLE_FACTORS M-Chon-08 M_TGFb_PA_2 Sample source:Mice Chondrocytes | Genotype:WT | Treatment:TGFb1 Label=U-13C-Palmitate; RAW_FILE_NAME(Raw file name)=M-Chon-08.mzML SUBJECT_SAMPLE_FACTORS M-Chon-09 M_TGFb_PA_3 Sample source:Mice Chondrocytes | Genotype:WT | Treatment:TGFb1 Label=U-13C-Palmitate; RAW_FILE_NAME(Raw file name)=M-Chon-09.mzML SUBJECT_SAMPLE_FACTORS M-Chon-10 M_BMP_PA_1 Sample source:Mice Chondrocytes | Genotype:WT | Treatment:BMP2 Label=U-13C-Palmitate; RAW_FILE_NAME(Raw file name)=M-Chon-10.mzML SUBJECT_SAMPLE_FACTORS M-Chon-11 M_BMP_PA_2 Sample source:Mice Chondrocytes | Genotype:WT | Treatment:BMP2 Label=U-13C-Palmitate; RAW_FILE_NAME(Raw file name)=M-Chon-11.mzML SUBJECT_SAMPLE_FACTORS M-Chon-12 M_BMP_PA_3 Sample source:Mice Chondrocytes | Genotype:WT | Treatment:BMP2 Label=U-13C-Palmitate; RAW_FILE_NAME(Raw file name)=M-Chon-12.mzML SUBJECT_SAMPLE_FACTORS M-Chon-13 M_TGFb_OA_1 Sample source:Mice Chondrocytes | Genotype:WT | Treatment:TGFb1 Label=U-13C-Oleate; RAW_FILE_NAME(Raw file name)=M-Chon-13.mzML SUBJECT_SAMPLE_FACTORS M-Chon-14 M_TGFb_OA_2 Sample source:Mice Chondrocytes | Genotype:WT | Treatment:TGFb1 Label=U-13C-Oleate; RAW_FILE_NAME(Raw file name)=M-Chon-14.mzML SUBJECT_SAMPLE_FACTORS M-Chon-15 M_TGFb_OA_3 Sample source:Mice Chondrocytes | Genotype:WT | Treatment:TGFb1 Label=U-13C-Oleate; RAW_FILE_NAME(Raw file name)=M-Chon-15.mzML SUBJECT_SAMPLE_FACTORS M-Chon-16 M_BMP_OA_1 Sample source:Mice Chondrocytes | Genotype:WT | Treatment:BMP2 Label=U-13C-Oleate; RAW_FILE_NAME(Raw file name)=M-Chon-16.mzML SUBJECT_SAMPLE_FACTORS M-Chon-17 M_BMP_OA_2 Sample source:Mice Chondrocytes | Genotype:WT | Treatment:BMP2 Label=U-13C-Oleate; RAW_FILE_NAME(Raw file name)=M-Chon-17.mzML SUBJECT_SAMPLE_FACTORS M-Chon-18 M_BMP_OA_3 Sample source:Mice Chondrocytes | Genotype:WT | Treatment:BMP2 Label=U-13C-Oleate; RAW_FILE_NAME(Raw file name)=M-Chon-18.mzML SUBJECT_SAMPLE_FACTORS H-Chon-01 H_Healthy_Gln_1 Sample source:Human Chondrocytes | Genotype:Healthy | Treatment:N/A Label=U-13C-Glutamine; RAW_FILE_NAME(Raw file name)=H-Chon-01.mzML SUBJECT_SAMPLE_FACTORS H-Chon-02 H_Healthy_Gln_2 Sample source:Human Chondrocytes | Genotype:Healthy | Treatment:N/A Label=U-13C-Glutamine; RAW_FILE_NAME(Raw file name)=H-Chon-02.mzML SUBJECT_SAMPLE_FACTORS H-Chon-03 H_Healthy_Gln_3 Sample source:Human Chondrocytes | Genotype:Healthy | Treatment:N/A Label=U-13C-Glutamine; RAW_FILE_NAME(Raw file name)=H-Chon-03.mzML SUBJECT_SAMPLE_FACTORS H-Chon-04 H_OA_Gln_1 Sample source:Human Chondrocytes | Genotype:OA | Treatment:N/A Label=U-13C-Glutamine; RAW_FILE_NAME(Raw file name)=H-Chon-04.mzML SUBJECT_SAMPLE_FACTORS H-Chon-05 H_OA_Gln_2 Sample source:Human Chondrocytes | Genotype:OA | Treatment:N/A Label=U-13C-Glutamine; RAW_FILE_NAME(Raw file name)=H-Chon-05.mzML SUBJECT_SAMPLE_FACTORS H-Chon-06 H_OA_Gln_3 Sample source:Human Chondrocytes | Genotype:OA | Treatment:N/A Label=U-13C-Glutamine; RAW_FILE_NAME(Raw file name)=H-Chon-06.mzML SUBJECT_SAMPLE_FACTORS H-Chon-07 H_Healthy_PA_1 Sample source:Human Chondrocytes | Genotype:Healthy | Treatment:N/A Label=U-13C-Palmitate; RAW_FILE_NAME(Raw file name)=H-Chon-07.mzML SUBJECT_SAMPLE_FACTORS H-Chon-08 H_Healthy_PA_2 Sample source:Human Chondrocytes | Genotype:Healthy | Treatment:N/A Label=U-13C-Palmitate; RAW_FILE_NAME(Raw file name)=H-Chon-08.mzML SUBJECT_SAMPLE_FACTORS H-Chon-09 H_Healthy_PA_3 Sample source:Human Chondrocytes | Genotype:Healthy | Treatment:N/A Label=U-13C-Palmitate; RAW_FILE_NAME(Raw file name)=H-Chon-09.mzML SUBJECT_SAMPLE_FACTORS H-Chon-10 H_OA_PA_1 Sample source:Human Chondrocytes | Genotype:OA | Treatment:N/A Label=U-13C-Palmitate; RAW_FILE_NAME(Raw file name)=H-Chon-10.mzML SUBJECT_SAMPLE_FACTORS H-Chon-11 H_OA_PA_2 Sample source:Human Chondrocytes | Genotype:OA | Treatment:N/A Label=U-13C-Palmitate; RAW_FILE_NAME(Raw file name)=H-Chon-11.mzML SUBJECT_SAMPLE_FACTORS H-Chon-12 H_OA_PA_3 Sample source:Human Chondrocytes | Genotype:OA | Treatment:N/A Label=U-13C-Palmitate; RAW_FILE_NAME(Raw file name)=H-Chon-12.mzML SUBJECT_SAMPLE_FACTORS H-Chon-13 H_Healthy_OA_1 Sample source:Human Chondrocytes | Genotype:Healthy | Treatment:N/A Label=U-13C-Oleate; RAW_FILE_NAME(Raw file name)=H-Chon-13.mzML SUBJECT_SAMPLE_FACTORS H-Chon-14 H_Healthy_OA_2 Sample source:Human Chondrocytes | Genotype:Healthy | Treatment:N/A Label=U-13C-Oleate; RAW_FILE_NAME(Raw file name)=H-Chon-14.mzML SUBJECT_SAMPLE_FACTORS H-Chon-15 H_Healthy_OA_3 Sample source:Human Chondrocytes | Genotype:Healthy | Treatment:N/A Label=U-13C-Oleate; RAW_FILE_NAME(Raw file name)=H-Chon-15.mzML SUBJECT_SAMPLE_FACTORS H-Chon-16 H_OA_OA_1 Sample source:Human Chondrocytes | Genotype:OA | Treatment:N/A Label=U-13C-Oleate; RAW_FILE_NAME(Raw file name)=H-Chon-16.mzML SUBJECT_SAMPLE_FACTORS H-Chon-17 H_OA_OA_2 Sample source:Human Chondrocytes | Genotype:OA | Treatment:N/A Label=U-13C-Oleate; RAW_FILE_NAME(Raw file name)=H-Chon-17.mzML SUBJECT_SAMPLE_FACTORS H-Chon-18 H_OA_OA_3 Sample source:Human Chondrocytes | Genotype:OA | Treatment:N/A Label=U-13C-Oleate; RAW_FILE_NAME(Raw file name)=H-Chon-18.mzML SUBJECT_SAMPLE_FACTORS H-Chon-19 H_Ctrl_Gln_1 Sample source:Human Chondrocytes | Genotype:OA | Treatment:Lenti-GFP Label=U-13C-Glutamine; RAW_FILE_NAME(Raw file name)=H-Chon-19.mzML SUBJECT_SAMPLE_FACTORS H-Chon-20 H_Ctrl_Gln_2 Sample source:Human Chondrocytes | Genotype:OA | Treatment:Lenti-GFP Label=U-13C-Glutamine; RAW_FILE_NAME(Raw file name)=H-Chon-20.mzML SUBJECT_SAMPLE_FACTORS H-Chon-21 H_Ctrl_Gln_3 Sample source:Human Chondrocytes | Genotype:OA | Treatment:Lenti-GFP Label=U-13C-Glutamine; RAW_FILE_NAME(Raw file name)=H-Chon-21.mzML SUBJECT_SAMPLE_FACTORS H-Chon-22 H_Lenti_Gln_1 Sample source:Human Chondrocytes | Genotype:OA | Treatment:Lenti-shNFIA Label=U-13C-Glutamine; RAW_FILE_NAME(Raw file name)=H-Chon-22.mzML SUBJECT_SAMPLE_FACTORS H-Chon-23 H_Lenti_Gln_2 Sample source:Human Chondrocytes | Genotype:OA | Treatment:Lenti-shNFIA Label=U-13C-Glutamine; RAW_FILE_NAME(Raw file name)=H-Chon-23.mzML SUBJECT_SAMPLE_FACTORS H-Chon-24 H_Lenti_Gln_3 Sample source:Human Chondrocytes | Genotype:OA | Treatment:Lenti-shNFIA Label=U-13C-Glutamine; RAW_FILE_NAME(Raw file name)=H-Chon-24.mzML SUBJECT_SAMPLE_FACTORS H-Chon-25 H_Ctrl_PA_1 Sample source:Human Chondrocytes | Genotype:OA | Treatment:Lenti-GFP Label=U-13C-Palmitate; RAW_FILE_NAME(Raw file name)=H-Chon-25.mzML SUBJECT_SAMPLE_FACTORS H-Chon-26 H_Ctrl_PA_2 Sample source:Human Chondrocytes | Genotype:OA | Treatment:Lenti-GFP Label=U-13C-Palmitate; RAW_FILE_NAME(Raw file name)=H-Chon-26.mzML SUBJECT_SAMPLE_FACTORS H-Chon-27 H_Ctrl_PA_3 Sample source:Human Chondrocytes | Genotype:OA | Treatment:Lenti-GFP Label=U-13C-Palmitate; RAW_FILE_NAME(Raw file name)=H-Chon-27.mzML SUBJECT_SAMPLE_FACTORS H-Chon-28 H_Lenti_PA_1 Sample source:Human Chondrocytes | Genotype:OA | Treatment:Lenti-shNFIA Label=U-13C-Palmitate; RAW_FILE_NAME(Raw file name)=H-Chon-28.mzML SUBJECT_SAMPLE_FACTORS H-Chon-29 H_Lenti_PA_2 Sample source:Human Chondrocytes | Genotype:OA | Treatment:Lenti-shNFIA Label=U-13C-Palmitate; RAW_FILE_NAME(Raw file name)=H-Chon-29.mzML SUBJECT_SAMPLE_FACTORS H-Chon-30 H_Lenti_PA_3 Sample source:Human Chondrocytes | Genotype:OA | Treatment:Lenti-shNFIA Label=U-13C-Palmitate; RAW_FILE_NAME(Raw file name)=H-Chon-30.mzML #COLLECTION CO:COLLECTION_SUMMARY Human articular chondrocytes were isolated from the femoral condyle and tibia CO:COLLECTION_SUMMARY plateau specimens obtained from healthy donors and OA patients. Following with CO:COLLECTION_SUMMARY 90 minutes digestion at 37 °C by 0.4% pronase and 14 hours digestion by 0.035% CO:COLLECTION_SUMMARY collagenase P, human articular chondrocytes were filtered through the 70 μm CO:COLLECTION_SUMMARY cell strainer and seeded at the density of 105 cells/cm2 for experiments. Murine CO:COLLECTION_SUMMARY articular chondrocytes were isolated from femoral heads of 3-week-old wildtype CO:COLLECTION_SUMMARY (C57BL/6J) pups. Following with 6 hours digestion at 37°C by 0.05% collagenase CO:COLLECTION_SUMMARY P, murine chondrocytes were harvested and plated at the density of 105 cells/cm2 CO:COLLECTION_SUMMARY for experiments. Both human and murine articular chondrocytes were cultured in CO:COLLECTION_SUMMARY the Dulbecco’s modified Eagle’s medium supplemented with 25 mM Glucose, 2 mM CO:COLLECTION_SUMMARY L-Glutamine, 10% fetal bovine serum, and 1% penicillin/streptomycin. The CO:COLLECTION_SUMMARY chondrocytes were treated with vehicle, TGF-β1 (5 ng/ml) and BMP2 (100 ng/ml) CO:COLLECTION_SUMMARY for 24-72 hours to examine the alterations of gene expression and metabolites. CO:COLLECTION_SUMMARY For NFIA/Nfia knockdown experiments, the chondrocytes were transfected with CO:COLLECTION_SUMMARY lentivirus expressing shRNA against NFIA and Nfia gene at a multiplicity of 10 CO:COLLECTION_SUMMARY in the presence of polybrene (10 μg/ml) in viral infection media (DMEM medium CO:COLLECTION_SUMMARY supplemented with 25 mM Glucose, 2 mM L-Glutamine and 2% FBS) for 24 hours. CO:COLLECTION_SUMMARY Lentivirus expressing shRNA against scramble sequences was used as control. For CO:COLLECTION_SUMMARY Nfia overexpression, the primary murine articular chondrocytes were transfected CO:COLLECTION_SUMMARY with lentivirus expressing murine Nfia gene at a multiplicity of 5 in the CO:COLLECTION_SUMMARY presence of polybrene for 48 hours. Lentivirus expression Gfp gene was used as CO:COLLECTION_SUMMARY control. CO:SAMPLE_TYPE Chondrocytes #TREATMENT TR:TREATMENT_SUMMARY The chondrocytes were treated with vehicle, TGF-β1 (5 ng/ml) and BMP2 (100 TR:TREATMENT_SUMMARY ng/ml). For isotope labeling, Articular chondrocytes were cultured in DMEM TR:TREATMENT_SUMMARY containing stable isotopically nutrient to assess the lipid metabolism in vitro. TR:TREATMENT_SUMMARY To assess lipids synthesis, articular chondrocytes were cultured in glutamine TR:TREATMENT_SUMMARY free DMEM supplemented with 10% dialyzed FBS, and 4 mM U-13C glutamine for 72 TR:TREATMENT_SUMMARY hours or cells were cultured in glucose free DMEM supplemented with 10% dialyzed TR:TREATMENT_SUMMARY FBS, and 25 mM U-13C glucose for 72 hours. To assess lipids oxidation, articular TR:TREATMENT_SUMMARY chondrocytes were cultured with 100 mM U-13C palmitic acid conjugated to BSA for TR:TREATMENT_SUMMARY 24 hours or 400 uM U-13C oleic acid. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Cell pellets were treated with a methanol/acetonitrile/water (2:2:1) solution SP:SAMPLEPREP_SUMMARY and reconstituted in 40 μL acetonitrile/water (1:1) per milligram dry weight. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE HILIC CH:INSTRUMENT_NAME Thermo Dionex Ultimate 3000 RS CH:COLUMN_NAME Merck SeQuant ZIC-pHILIC (100 x 2.1mm,5um) CH:SOLVENT_A 95% water/5% acetonitrile; 10 mM ammonium bicarbonate; 0.1% ammonium hydroxide; CH:SOLVENT_A 2.5 mM medronic acid CH:SOLVENT_B 95% acetonitrile/5% water; 2.5 uM medronic acid CH:FLOW_GRADIENT 90% B from 0-1 min and 90% to 25% B from 1–15 min CH:FLOW_RATE 250 uL/min CH:COLUMN_TEMPERATURE 40 #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Thermo Q Exactive Plus Orbitrap MS:INSTRUMENT_TYPE Orbitrap MS:MS_TYPE ESI MS:ION_MODE NEGATIVE MS:MS_COMMENTS Data were collected with the following MS source settings: spray voltage, - MS:MS_COMMENTS 2.5kV; sheath gas, 45; auxiliary gas, 10; sweep gas, 0; capillary temperature, MS:MS_COMMENTS 250°C; resolution, 140,000; maximum injection time, 100 ms. Skyline software MS:MS_COMMENTS was used for data processing #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS peak area MS_METABOLITE_DATA_START Samples H_Healthy_PA_1 H_Healthy_PA_2 H_Healthy_PA_3 H_OA_PA_1 H_OA_PA_2 H_OA_PA_3 H_Ctrl_PA_1 H_Ctrl_PA_2 H_Ctrl_PA_3 H_Lenti_PA_1 H_Lenti_PA_2 H_Lenti_PA_3 Factors Sample source:Human Chondrocytes | Genotype:Healthy | Treatment:N/A Sample source:Human Chondrocytes | Genotype:Healthy | Treatment:N/A Sample source:Human Chondrocytes | Genotype:Healthy | Treatment:N/A Sample source:Human Chondrocytes | Genotype:OA | Treatment:N/A Sample source:Human Chondrocytes | Genotype:OA | Treatment:N/A Sample source:Human Chondrocytes | Genotype:OA | Treatment:N/A Sample source:Human Chondrocytes | Genotype:OA | Treatment:Lenti-GFP Sample source:Human Chondrocytes | Genotype:OA | Treatment:Lenti-GFP Sample source:Human Chondrocytes | Genotype:OA | Treatment:Lenti-GFP Sample source:Human Chondrocytes | Genotype:OA | Treatment:Lenti-shNFIA Sample source:Human Chondrocytes | Genotype:OA | Treatment:Lenti-shNFIA Sample source:Human Chondrocytes | Genotype:OA | Treatment:Lenti-shNFIA Citrate 1191891279 1194003161 1102319706 2190240890 2055612951 2656871643 3160340403 3370171422 4021460897 2229815514 2495484260 2967070029 Fumarate 90934231.03 90404528.73 95915299.79 71812702.73 61411736.73 103888950 227239127.9 224363433.5 119381431 112635065.3 119953092.2 165449770.6 Malate 1601121878 1674196710 1789508952 1067978812 1119733605 1427198710 4409370374 4501364002 2497383288 2210385123 2364996497 3254667379 Glutamate 10221502096 11885974244 12613134296 17154527129 15409488225 20658127744 6611962675 7085794396 10313567239 4587475447 4374658062 4452225192 Aspartate 294100128.5 321262784.7 321027240 1800935634 1723692875 2001860654 667575823.3 621632494.5 605775992.3 688904974.8 599481169.2 996797436.3 ATP 205903468.5 241522123.6 265863680.4 347357635.7 338339737.4 449409123.9 345139167.5 310113018.3 266706163.6 190554624.9 212028124.5 239070486.9 NAD+ 211661836.3 236211458.4 245357582.6 128673376.2 118026049 151480203.6 285708615.6 287673471.5 206143413.5 140562982.9 146320637.7 162397167.8 NADH 7686087.849 11780798.06 7558748.84 1700038.012 1636801.646 2951680.396 24547421.34 16867942.08 44607110.32 4758405.653 1162166.844 4342732.994 dGTP 205159001.6 240504382.3 264855395.4 346330843.5 337329319 447985885.8 349363842.3 313887409.7 270511992.4 193096184.3 214304534.4 242276803.1 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name retention index quantified m/z KEGG ID Citrate 8.4 191 C00158 Fumarate 7 115 C00122 Malate 7 133 C00497 Glutamate 6 146 C00025 Aspartate 6.2 132 C16433 ATP 7.8 506 C00002 NAD+ 6 662 C00003 NADH 5.8 664 C00004 dGTP 7.8 506 C00286 METABOLITES_END #END