#METABOLOMICS WORKBENCH kevin.cho@wustl.edu_20241126_081352 DATATRACK_ID:5407 STUDY_ID:ST003598 ANALYSIS_ID:AN005913 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 Reversed phase CH:INSTRUMENT_NAME Agilent 1290 Infinity CH:COLUMN_NAME Phenomenex Kinetex C18 (150 x 2.1mm,2.6um) CH:SOLVENT_A 100% water; 0.1% formic acid CH:SOLVENT_B 100% acetonitrile; 0.1% formic acid CH:FLOW_GRADIENT 30% B from 0-3 min, 30% B to 95% B from 3–30 min and 95% B from 30–40 min CH:FLOW_RATE 200 uL/min CH:COLUMN_TEMPERATURE 40 #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Agilent 6540 QTOF MS:INSTRUMENT_TYPE QTOF MS:MS_TYPE ESI MS:ION_MODE POSITIVE MS:MS_COMMENTS Data were collected with the following MS source settings: gas temperature, MS:MS_COMMENTS 300°C; gas flow, 12 l/min; sheath gas temperature, 300°C; sheath gas flow, 11 MS:MS_COMMENTS l/min; nebulizer, 30 psi; Vcap, 3kV; nozzle voltage, 1 kV. Skyline software was MS:MS_COMMENTS used for data processing. #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS peak area MS_METABOLITE_DATA_START Samples M_TGFb_Gln_1 M_TGFb_Gln_2 M_TGFb_Gln_3 M_BMP_Gln_1 M_BMP_Gln_2 M_BMP_Gln_3 H_Healthy_Gln_1 H_Healthy_Gln_2 H_Healthy_Gln_3 H_OA_Gln_1 H_OA_Gln_2 H_OA_Gln_3 H_Ctrl_Gln_1 H_Ctrl_Gln_2 H_Ctrl_Gln_3 H_Lenti_Gln_1 H_Lenti_Gln_2 H_Lenti_Gln_3 Factors Sample source:Mice Chondrocytes | Genotype:WT | Treatment:TGFb1 Sample source:Mice Chondrocytes | Genotype:WT | Treatment:TGFb1 Sample source:Mice Chondrocytes | Genotype:WT | Treatment:TGFb1 Sample source:Mice Chondrocytes | Genotype:WT | Treatment:BMP2 Sample source:Mice Chondrocytes | Genotype:WT | Treatment:BMP2 Sample source:Mice Chondrocytes | Genotype:WT | Treatment:BMP2 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 CAR 16:0 69826.73128 81288.27194 147689.0602 3681896.716 3268476.7 3101321.035 144312.0906 326122.5253 285392.1021 264366.867 150425.2776 66532.35381 958197.1544 2340559.399 5225255.086 3594907.226 3201897.334 3034900.414 CAR 18:0 21537.99258 17126.32015 35003.51044 1966548.088 1744506.579 1674314.169 36252.95411 68002.26474 71805.30542 114187.8739 58766.70131 30095.87671 563147.7251 1254340.918 3188171 1941280.291 1741546.417 1667475.324 CAR 18:1 94287.48218 109726.5876 244085.2544 2903930.522 2643823.712 2507908.215 206908.8404 524881.7161 421458.6348 399639.9428 202151.7755 77963.77299 522091.8617 1688025.03 4880555.017 2840292.107 2566821.18 2439469.745 LPC 16:0 569995.5415 687920.2441 963363.1783 1688088.565 1396757.375 1458764.678 693671.4367 1447576.442 1237996.584 3217879.818 2027371.267 919237.2362 484867.0724 970218.2218 2228463.525 1741914.522 1439845.348 1516104.886 LPC 18:0 463039.6775 474675.6792 585664.8822 1311416.714 1207018.85 1205983.534 439436.9615 869888.6991 871705.062 2710647.767 1942664.101 886770.592 391100.9392 686864.7126 1596166.661 1322206.806 1209056.113 1243079.402 LPE 16:0 15205.03048 17765.33843 14063.79151 123107.5651 112848.1754 118516.1017 12091.231 24361.14235 17728.13129 100442.6183 77400.32154 43136.55019 44684.12469 61475.15714 111220.1058 122275.9421 111379.5271 117525.1664 LPE 18:0 46864.33262 43832.64939 24547.0418 213824.5141 224361.8915 224583.4164 27151.77112 58091.40745 49835.69911 183151.0089 132664.228 87540.59292 146042.733 191914.0051 363612.4796 210851.8683 221125.2755 223454.1794 PC 16:0_18:1 786107.4851 27377686.43 17302535.13 19647620.99 8003966.812 6303666.672 227559.0525 1900168.695 1054553.131 3139626.492 6866915.623 6762272.995 11007969.47 10634792.54 5021188.223 22954983.72 8510244.055 6855758.067 PC 36:3 2675552.583 1565191.245 945278.6272 2162222.388 2007346.826 2002385.659 136000.0111 229087.5816 144135.0263 2433058.953 1696317.94 358329.1169 539756.05 674178.3584 2136775.323 1959831.302 1880257.543 1815318.649 PC 36:4 7893381.363 4538899.602 2683055.177 17667603.82 15936574.25 15485716.44 496819.9736 861890.1054 564223.1774 6419755.666 4393415.056 1009660.594 3313159.521 4132725.377 12528868.39 15502886.8 14843490.59 13918618.85 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name retention time quantified m/z KEGG ID CAR 16:0 14.4 400 C02990 CAR 18:0 16.6 428 CAR 18:1 14.9 426 LPC 16:0 15.9 496 LPC 18:0 18.8 524 LPE 16:0 15.7 454 LPE 18:0 18.7 482 PC 16:0_18:1 30.5 760 PC 36:3 37.7 785 PC 36:4 37.9 783 METABOLITES_END #END