#METABOLOMICS WORKBENCH wongw24_20241114_172657 DATATRACK_ID:5371 STUDY_ID:ST003578 ANALYSIS_ID:AN005875 PROJECT_ID:PR002208 VERSION 1 CREATED_ON 12-06-2024 #PROJECT PR:PROJECT_TITLE NRF2 supports non-small cell lung cancer growth independently of PR:PROJECT_TITLE CBP/p300-enhanced glutathione synthesis PR:PROJECT_TYPE MS quantitative analysis PR:PROJECT_SUMMARY Nuclear factor erythroid 2-related factor 2 (NRF2) is a stress responsive PR:PROJECT_SUMMARY transcription factor that is mutationally activated in a subset (~25%) of PR:PROJECT_SUMMARY clinically-aggressive non-small cell lung cancers (NSCLC). Mechanistic insight PR:PROJECT_SUMMARY into drivers of the NRF2 dependency remains poorly understood. Here, we defined PR:PROJECT_SUMMARY a novel NRF2 target gene set linked to NRF2-dependency in cancer cell lines, and PR:PROJECT_SUMMARY observed that a significant portion of these genes is devoid of PR:PROJECT_SUMMARY promoter-proximal NRF2. Using integrated genomic analyses, we characterized PR:PROJECT_SUMMARY extensive NRF2-dependent enhancer RNA (eRNA) synthesis and NRF2-mediated H3K27ac PR:PROJECT_SUMMARY deposition at proximal and distal enhancer regions regulating these genes. While PR:PROJECT_SUMMARY CBP/p300 is a well-validated direct interaction partner of NRF2 with prominent PR:PROJECT_SUMMARY functions at enhancers, we report that this interaction is not required for PR:PROJECT_SUMMARY NRF2-dependent NSCLC cell growth, indicating that NRF2 can sustain sufficient PR:PROJECT_SUMMARY transcriptional activity in the absence of CBP/p300 coactivation. Broad PR:PROJECT_SUMMARY metabolic profiling established a primary role for CBP/p300 in NRF2-dependent PR:PROJECT_SUMMARY accumulation of glutathione and glutathione-related metabolites. While redox PR:PROJECT_SUMMARY homeostasis via enhanced glutathione production is commonly associated with the PR:PROJECT_SUMMARY normal physiological role of NRF2, collectively our results suggest that PR:PROJECT_SUMMARY NRF2-dependent cancer cell growth does not require this enhanced glutathione PR:PROJECT_SUMMARY production. PR:INSTITUTE Genentech Inc. PR:LAST_NAME Wong PR:FIRST_NAME Weng Ruh PR:ADDRESS 1 DNA Way, South San Francisco, CA 94080, USA PR:EMAIL wongw24@gene.com PR:PHONE 4089048962 PR:DOI http://dx.doi.org/10.21228/M86Z6P PR:CONTRIBUTORS Ryan J. Conrad, James A. Mondo, Mike Lingjue Wang, Peter S. Liu, Zijuan Lai, PR:CONTRIBUTORS Feroza K Choudhury, Qingling Li, Weng Ruh Wong, James Lee, Frances Shanahan, Eva PR:CONTRIBUTORS Lin, Scott Martin, Joachim Rudolph, John G. Moffat, Dewakar Sangaraju, Wendy PR:CONTRIBUTORS Sandoval, Timothy Sterne-Weiler, Scott A. Foster #STUDY ST:STUDY_TITLE NRF2 supports non-small cell lung cancer growth independently of ST:STUDY_TITLE CBP/p300-enhanced glutathione synthesis ST:STUDY_TYPE MS quantitative analysis ST:STUDY_SUMMARY Nuclear factor erythroid 2-related factor 2 (NRF2) is a stress responsive ST:STUDY_SUMMARY transcription factor that is mutationally activated in a subset (~25%) of ST:STUDY_SUMMARY clinically-aggressive non-small cell lung cancers (NSCLC). Mechanistic insight ST:STUDY_SUMMARY into drivers of the NRF2 dependency remains poorly understood. Here, we defined ST:STUDY_SUMMARY a novel NRF2 target gene set linked to NRF2-dependency in cancer cell lines, and ST:STUDY_SUMMARY observed that a significant portion of these genes is devoid of ST:STUDY_SUMMARY promoter-proximal NRF2. Using integrated genomic analyses, we characterized ST:STUDY_SUMMARY extensive NRF2-dependent enhancer RNA (eRNA) synthesis and NRF2-mediated H3K27ac ST:STUDY_SUMMARY deposition at proximal and distal enhancer regions regulating these genes. While ST:STUDY_SUMMARY CBP/p300 is a well-validated direct interaction partner of NRF2 with prominent ST:STUDY_SUMMARY functions at enhancers, we report that this interaction is not required for ST:STUDY_SUMMARY NRF2-dependent NSCLC cell growth, indicating that NRF2 can sustain sufficient ST:STUDY_SUMMARY transcriptional activity in the absence of CBP/p300 coactivation. Broad ST:STUDY_SUMMARY metabolic profiling established a primary role for CBP/p300 in NRF2-dependent ST:STUDY_SUMMARY accumulation of glutathione and glutathione-related metabolites. While redox ST:STUDY_SUMMARY homeostasis via enhanced glutathione production is commonly associated with the ST:STUDY_SUMMARY normal physiological role of NRF2, collectively our results suggest that ST:STUDY_SUMMARY NRF2-dependent cancer cell growth does not require this enhanced glutathione ST:STUDY_SUMMARY production. ST:INSTITUTE Genentech Inc. ST:LAST_NAME Wong ST:FIRST_NAME Weng Ruh ST:ADDRESS 1 DNA Way, South San Francisco, CA 94080, USA ST:EMAIL wongw24@gene.com ST:PHONE 4089048962 ST:SUBMIT_DATE 2024-11-14 #SUBJECT SU:SUBJECT_TYPE Cultured cells SU:SUBJECT_SPECIES Homo sapiens SU:TAXONOMY_ID 9606 SU:GENOTYPE_STRAIN A549 SU:SPECIES_GROUP Mammals #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data SUBJECT_SAMPLE_FACTORS - luc+1 Sample source:A549 human lung epithelial cells | Condition:Luciferase + Dox RAW_FILE_NAME_1=Lipidyzer Batch - 20211014120816 - 1-30545 - luc+1.mzML; RAW_FILE_NAME_2=Lipidyzer Batch - 20211014120816 - 2-30594 - luc+1.mzML SUBJECT_SAMPLE_FACTORS - luc+2 Sample source:A549 human lung epithelial cells | Condition:Luciferase + Dox RAW_FILE_NAME_1=Lipidyzer Batch - 20211014120816 - 1-30546 - luc+2.mzML; RAW_FILE_NAME_2=Lipidyzer Batch - 20211014120816 - 2-30595 - luc+2.mzML SUBJECT_SAMPLE_FACTORS - luc+3 Sample source:A549 human lung epithelial cells | Condition:Luciferase + Dox RAW_FILE_NAME_1=Lipidyzer Batch - 20211014120816 - 1-30547 - luc+3.mzML; RAW_FILE_NAME_2=Lipidyzer Batch - 20211014120816 - 2-30596 - luc+3.mzML SUBJECT_SAMPLE_FACTORS - luc+4 Sample source:A549 human lung epithelial cells | Condition:Luciferase + Dox RAW_FILE_NAME_1=Lipidyzer Batch - 20211014120816 - 1-30548 - luc+4.mzML; RAW_FILE_NAME_2=Lipidyzer Batch - 20211014120816 - 2-30597 - luc+4.mzML SUBJECT_SAMPLE_FACTORS - luc-1 Sample source:A549 human lung epithelial cells | Condition:Luciferase - Dox RAW_FILE_NAME_1=Lipidyzer Batch - 20211014120816 - 1-30529 - luc-1.mzML; RAW_FILE_NAME_2=Lipidyzer Batch - 20211014120816 - 2-30578 - luc-1.mzML SUBJECT_SAMPLE_FACTORS - luc-2 Sample source:A549 human lung epithelial cells | Condition:Luciferase - Dox RAW_FILE_NAME_1=Lipidyzer Batch - 20211014120816 - 1-30530 - luc-2.mzML; RAW_FILE_NAME_2=Lipidyzer Batch - 20211014120816 - 2-30579 - luc-2.mzML SUBJECT_SAMPLE_FACTORS - luc-3 Sample source:A549 human lung epithelial cells | Condition:Luciferase - Dox RAW_FILE_NAME_1=Lipidyzer Batch - 20211014120816 - 1-30531 - luc-3.mzML; RAW_FILE_NAME_2=Lipidyzer Batch - 20211014120816 - 2-30580 - luc-3.mzML SUBJECT_SAMPLE_FACTORS - luc-4 Sample source:A549 human lung epithelial cells | Condition:Luciferase - Dox RAW_FILE_NAME_1=Lipidyzer Batch - 20211014120816 - 1-30532 - luc-4.mzML; RAW_FILE_NAME_2=Lipidyzer Batch - 20211014120816 - 2-30581 - luc-4.mzML SUBJECT_SAMPLE_FACTORS - FL+1 Sample source:A549 human lung epithelial cells | Condition:NRF2 Full-length + Dox RAW_FILE_NAME_1=Lipidyzer Batch - 20211014120816 - 1-30549 - FL+1.mzML; RAW_FILE_NAME_2=Lipidyzer Batch - 20211014120816 - 2-30598 - FL+1.mzML SUBJECT_SAMPLE_FACTORS - FL+2 Sample source:A549 human lung epithelial cells | Condition:NRF2 Full-length + Dox RAW_FILE_NAME_1=Lipidyzer Batch - 20211014120816 - 1-30550 - FL+2.mzML; RAW_FILE_NAME_2=Lipidyzer Batch - 20211014120816 - 2-30599 - FL+2.mzML SUBJECT_SAMPLE_FACTORS - FL+3 Sample source:A549 human lung epithelial cells | Condition:NRF2 Full-length + Dox RAW_FILE_NAME_1=Lipidyzer Batch - 20211014120816 - 1-30551 - FL+3.mzML; RAW_FILE_NAME_2=Lipidyzer Batch - 20211014120816 - 2-30600 - FL+3.mzML SUBJECT_SAMPLE_FACTORS - FL+4 Sample source:A549 human lung epithelial cells | Condition:NRF2 Full-length + Dox RAW_FILE_NAME_1=Lipidyzer Batch - 20211014120816 - 1-30552 - FL+4.mzML; RAW_FILE_NAME_2=Lipidyzer Batch - 20211014120816 - 2-30601 - FL+4.mzML SUBJECT_SAMPLE_FACTORS - FL-1 Sample source:A549 human lung epithelial cells | Condition:NRF2 Full-length - Dox RAW_FILE_NAME_1=Lipidyzer Batch - 20211014120816 - 1-30533 - FL-1.mzML; RAW_FILE_NAME_2=Lipidyzer Batch - 20211014120816 - 2-30582 - FL-1.mzML SUBJECT_SAMPLE_FACTORS - FL-2 Sample source:A549 human lung epithelial cells | Condition:NRF2 Full-length - Dox RAW_FILE_NAME_1=Lipidyzer Batch - 20211014120816 - 1-30534 - FL-2.mzML; RAW_FILE_NAME_2=Lipidyzer Batch - 20211014120816 - 2-30583 - FL-2.mzML SUBJECT_SAMPLE_FACTORS - FL-3 Sample source:A549 human lung epithelial cells | Condition:NRF2 Full-length - Dox RAW_FILE_NAME_1=Lipidyzer Batch - 20211014120816 - 1-30535 - FL-3.mzML; RAW_FILE_NAME_2=Lipidyzer Batch - 20211014120816 - 2-30584 - FL-3.mzML SUBJECT_SAMPLE_FACTORS - FL-4 Sample source:A549 human lung epithelial cells | Condition:NRF2 Full-length - Dox RAW_FILE_NAME_1=Lipidyzer Batch - 20211014120816 - 1-30536 - FL-4.mzML; RAW_FILE_NAME_2=Lipidyzer Batch - 20211014120816 - 2-30585 - FL-4.mzML SUBJECT_SAMPLE_FACTORS - 45+1 Sample source:A549 human lung epithelial cells | Condition:NRF2 Neh4,5 mut + Dox RAW_FILE_NAME_1=Lipidyzer Batch - 20211014120816 - 1-30553 - 45+1.mzML; RAW_FILE_NAME_2=Lipidyzer Batch - 20211014120816 - 2-30602 - 45+1.mzML SUBJECT_SAMPLE_FACTORS - 45+2 Sample source:A549 human lung epithelial cells | Condition:NRF2 Neh4,5 mut + Dox RAW_FILE_NAME_1=Lipidyzer Batch - 20211014120816 - 1-30554 - 45+2.mzML; RAW_FILE_NAME_2=Lipidyzer Batch - 20211014120816 - 2-30603 - 45+2.mzML SUBJECT_SAMPLE_FACTORS - 45+3 Sample source:A549 human lung epithelial cells | Condition:NRF2 Neh4,5 mut + Dox RAW_FILE_NAME_1=Lipidyzer Batch - 20211014120816 - 1-30555 - 45+3.mzML; RAW_FILE_NAME_2=Lipidyzer Batch - 20211014120816 - 2-30604 - 45+3.mzML SUBJECT_SAMPLE_FACTORS - 45+4 Sample source:A549 human lung epithelial cells | Condition:NRF2 Neh4,5 mut + Dox RAW_FILE_NAME_1=Lipidyzer Batch - 20211014120816 - 1-30556 - 45+4.mzML; RAW_FILE_NAME_2=Lipidyzer Batch - 20211014120816 - 2-30605 - 45+4.mzML SUBJECT_SAMPLE_FACTORS - 45-1 Sample source:A549 human lung epithelial cells | Condition:NRF2 Neh4,5 mut - Dox RAW_FILE_NAME_1=Lipidyzer Batch - 20211014120816 - 1-30537 - 45-1.mzML; RAW_FILE_NAME_2=Lipidyzer Batch - 20211014120816 - 2-30586 - 45-1.mzML SUBJECT_SAMPLE_FACTORS - 45-2 Sample source:A549 human lung epithelial cells | Condition:NRF2 Neh4,5 mut - Dox RAW_FILE_NAME_1=Lipidyzer Batch - 20211014120816 - 1-30538 - 45-2.mzML; RAW_FILE_NAME_2=Lipidyzer Batch - 20211014120816 - 2-30587 - 45-2.mzML SUBJECT_SAMPLE_FACTORS - 45-3 Sample source:A549 human lung epithelial cells | Condition:NRF2 Neh4,5 mut - Dox RAW_FILE_NAME_1=Lipidyzer Batch - 20211014120816 - 1-30539 - 45-3.mzML; RAW_FILE_NAME_2=Lipidyzer Batch - 20211014120816 - 2-30588 - 45-3.mzML SUBJECT_SAMPLE_FACTORS - 45-4 Sample source:A549 human lung epithelial cells | Condition:NRF2 Neh4,5 mut - Dox RAW_FILE_NAME_1=Lipidyzer Batch - 20211014120816 - 1-30540 - 45-4.mzML; RAW_FILE_NAME_2=Lipidyzer Batch - 20211014120816 - 2-30589 - 45-4.mzML #COLLECTION CO:COLLECTION_SUMMARY All cell lines were obtained from the Genentech cell bank (Yu et al, 2015) and CO:COLLECTION_SUMMARY grown in RPMI or DMEM supplemented with 10% FBS, 1% glutamine, and 1% CO:COLLECTION_SUMMARY penicillin-streptomycin at 37°C, 5% CO2. Cell lines were confirmed via STR CO:COLLECTION_SUMMARY profiling and were mycoplasma free. A549 and A549shNRF2-BIND rescue cell lines CO:COLLECTION_SUMMARY were generated using the Piggybac system. Donor constructs were co-transfected CO:COLLECTION_SUMMARY with transposase (Ding et al, 2005) at a 4:1 ratio using Lipofetamine 3000 CO:COLLECTION_SUMMARY (Thermo) according to manufacturerâs protocol. 3-days post-transfection, CO:COLLECTION_SUMMARY puromycin selection (1µg/mL) was initiated and was performed for ~2 weeks prior CO:COLLECTION_SUMMARY to experimentation. A549shNRF2-BIND rescue cells represent polyclonal pools. CO:COLLECTION_SUMMARY Cell viability was measured using CellTiter-Glo® Luminescent Cell Viability CO:COLLECTION_SUMMARY Assay (Promega) using the Envision 2103 Plate Reader (Promega). Cell growth CO:COLLECTION_SUMMARY curves were generated using the IncuCyte System (Sartorius) according to the CO:COLLECTION_SUMMARY manufacturerâs protocol. Curve fits were performed in Prism. Clonogenic assays CO:COLLECTION_SUMMARY were performed by washing cells in PBS, staining with 0.5% crystal violet CO:COLLECTION_SUMMARY solution (Sigma HT90132) for 5 min, and washing 3x with PBS. Four replicates of CO:COLLECTION_SUMMARY 20-30 million A549shNRF2-BIND-luc, WT NRF2 or Neh4/5mut cells were treated with CO:COLLECTION_SUMMARY 2.5ng/mL dox for 48h, upon which cells were trypsinized, washed 3x with ice cold CO:COLLECTION_SUMMARY PBS and flash frozen in liquid nitrogen for metabolic quenching. CO:COLLECTION_PROTOCOL_FILENAME OBJ0041730_NRF2_lipidomics_protocol.pdf CO:SAMPLE_TYPE Epithelial cells #TREATMENT TR:TREATMENT_SUMMARY No treatment was administered and this is a study based on genetic variants. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY 500 ul of cold methanol was added to cell pellets. After ultrasonication for 30 SP:SAMPLEPREP_SUMMARY sec, cell lysate was transferred to extraction tubes. 500μL of water (HPLC SP:SAMPLEPREP_SUMMARY grade, Fisher Chemical), 500μL cold methanol (HPLC grade, Fisher Chemical) and SP:SAMPLEPREP_SUMMARY 450μL Dichloromethane (HPLC grade, Fisher Chemical) were added to the cell SP:SAMPLEPREP_SUMMARY lysate to form a single phase. Samples were incubated on ice for 30min. After SP:SAMPLEPREP_SUMMARY 30min, 100μL Lipidyzer internal standard (Sciex) was added to the sample SP:SAMPLEPREP_SUMMARY mixture, followed by 500μL of H2O and 450μL of Dichloromethane. Samples were SP:SAMPLEPREP_SUMMARY centrifuged at 1500rpm for 15min. Bottom layer was transferred to a new SP:SAMPLEPREP_SUMMARY collection tube. 0.9mL of Dichloromethane was then added to the extraction tube SP:SAMPLEPREP_SUMMARY for the second extraction. The bottom layer was combined to the same collection SP:SAMPLEPREP_SUMMARY tube. Sample extract was then dried under gentle stream of nitrogen and SP:SAMPLEPREP_SUMMARY reconstituted in 500μL buffer (Dichloromethane:Methanol (1:1), 10mM ammonium SP:SAMPLEPREP_SUMMARY acetate) for lipidomics analysis #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY Differential Mobility Spectrometry turned on with switching pos/neg polarity CH:CHROMATOGRAPHY_COMMENTS Reference: Ubhi, B.K. (2018). Direct Infusion-Tandem Mass Spectrometry CH:CHROMATOGRAPHY_COMMENTS (DI-MS/MS) Analysis of Complex Lipids in Human Plasma and Serum Using the CH:CHROMATOGRAPHY_COMMENTS Lipidyzer⢠Platform. In: Giera, M. (eds) Clinical Metabolomics. Methods in CH:CHROMATOGRAPHY_COMMENTS Molecular Biology, vol 1730. Humana Press, New York, NY. CH:CHROMATOGRAPHY_COMMENTS https://doi.org/10.1007/978-1-4939-7592-1_15 CH:INSTRUMENT_NAME Sciex QTRAP 6500+ with SelexION CH:COLUMN_NAME none CH:COLUMN_TEMPERATURE none CH:FLOW_GRADIENT none CH:FLOW_RATE none CH:SAMPLE_INJECTION 50 ul CH:SOLVENT_A none CH:SOLVENT_B none CH:CHROMATOGRAPHY_TYPE Flow injection analysis #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME ABI Sciex 6500+ MS:INSTRUMENT_TYPE QTRAP MS:MS_TYPE ESI MS:MS_COMMENTS DI-MS/MS Analysis 1. A QTRAP® system with SelexION Technology (SCIEX) is used MS:MS_COMMENTS for targeted profiling (SCIEX, MA, USA). 2. This method is using a flow MS:MS_COMMENTS injection analysis (FIA): one injection with the SelexION voltages turned ON 3. MS:MS_COMMENTS The lipid molecular species are measured using multiple reaction monitoring MS:MS_COMMENTS (MRM) and positive/negative switching. The Positive ion mode detected the MS:MS_COMMENTS following lipid classes: SM 4. A flow injection analysis (FIA) setup is employed MS:MS_COMMENTS by using the LC to flow at an isocratic rate of 7 μL/min with a ramp up to 30 MS:MS_COMMENTS μL/min for the last 2 min of the experiment to allow for washing. 5. Data MS:MS_COMMENTS acquisition is around 20 min per sample, and 50 μL of the reconstituted sample MS:MS_COMMENTS is infused and the area under the flat infusion line reported and corrected to MS:MS_COMMENTS the appropriate internal standard . 6. Samples are quantified using the LWM MS:MS_COMMENTS software which reports all the detected lipids. Refence: Ubhi, B.K. (2018). MS:MS_COMMENTS Direct Infusion-Tandem Mass Spectrometry (DI-MS/MS) Analysis of Complex Lipids MS:MS_COMMENTS in Human Plasma and Serum Using the Lipidyzer⢠Platform. In: Giera, M. (eds) MS:MS_COMMENTS Clinical Metabolomics. Methods in Molecular Biology, vol 1730. Humana Press, New MS:MS_COMMENTS York, NY. https://doi.org/10.1007/978-1-4939-7592-1_15 MS:ION_MODE POSITIVE #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS nmol/million cells MS_METABOLITE_DATA_START Samples luc+1 luc+2 luc+3 luc+4 luc-1 luc-2 luc-3 luc-4 FL+1 FL+2 FL+3 FL+4 FL-1 FL-2 FL-3 FL-4 45+1 45+2 45+3 45+4 45-1 45-2 45-3 45-4 Factors Sample source:A549 human lung epithelial cells | Condition:Luciferase + Dox Sample source:A549 human lung epithelial cells | Condition:Luciferase + Dox Sample source:A549 human lung epithelial cells | Condition:Luciferase + Dox Sample source:A549 human lung epithelial cells | Condition:Luciferase + Dox Sample source:A549 human lung epithelial cells | Condition:Luciferase - Dox Sample source:A549 human lung epithelial cells | Condition:Luciferase - Dox Sample source:A549 human lung epithelial cells | Condition:Luciferase - Dox Sample source:A549 human lung epithelial cells | Condition:Luciferase - Dox Sample source:A549 human lung epithelial cells | Condition:NRF2 Full-length + Dox Sample source:A549 human lung epithelial cells | Condition:NRF2 Full-length + Dox Sample source:A549 human lung epithelial cells | Condition:NRF2 Full-length + Dox Sample source:A549 human lung epithelial cells | Condition:NRF2 Full-length + Dox Sample source:A549 human lung epithelial cells | Condition:NRF2 Full-length - Dox Sample source:A549 human lung epithelial cells | Condition:NRF2 Full-length - Dox Sample source:A549 human lung epithelial cells | Condition:NRF2 Full-length - Dox Sample source:A549 human lung epithelial cells | Condition:NRF2 Full-length - Dox Sample source:A549 human lung epithelial cells | Condition:NRF2 Neh4,5 mut + Dox Sample source:A549 human lung epithelial cells | Condition:NRF2 Neh4,5 mut + Dox Sample source:A549 human lung epithelial cells | Condition:NRF2 Neh4,5 mut + Dox Sample source:A549 human lung epithelial cells | Condition:NRF2 Neh4,5 mut + Dox Sample source:A549 human lung epithelial cells | Condition:NRF2 Neh4,5 mut - Dox Sample source:A549 human lung epithelial cells | Condition:NRF2 Neh4,5 mut - Dox Sample source:A549 human lung epithelial cells | Condition:NRF2 Neh4,5 mut - Dox Sample source:A549 human lung epithelial cells | Condition:NRF2 Neh4,5 mut - Dox SM(14:0) 0.1164 0.1120 0.1067 0.0957 0.0671 0.0673 0.0658 0.0789 0.0740 0.0748 0.0832 0.0839 0.0746 0.0744 0.0752 0.0855 0.0724 0.0735 0.0795 0.0830 0.0718 0.0649 0.0709 0.0676 SM(16:0) 2.2249 2.2081 2.1094 1.9232 1.5777 1.5271 1.4974 1.7487 1.7490 1.7523 1.8620 1.8938 1.7463 1.7376 1.7426 2.0521 1.3917 1.4540 1.5381 1.6342 1.4979 1.3954 1.5050 1.4333 SM(18:0) 0.1151 0.1149 0.1081 0.1035 0.0871 0.0833 0.0802 0.0918 0.0957 0.1011 0.1085 0.1057 0.0898 0.0865 0.0886 0.1020 0.0804 0.0901 0.0948 0.0978 0.0864 0.0797 0.0852 0.0814 SM(18:1) 0.0147 0.0146 0.0146 0.0124 0.0112 0.0103 0.0111 0.0124 0.0193 0.0189 0.0216 0.0189 0.0121 0.0118 0.0124 0.0126 0.0130 0.0138 0.0135 0.0147 0.0113 0.0112 0.0129 0.0113 SM(20:0) 0.1743 0.1683 0.1583 0.1546 0.1721 0.1532 0.1525 0.1674 0.1553 0.1600 0.1796 0.1640 0.1525 0.1434 0.1582 0.1675 0.1348 0.1500 0.1452 0.1713 0.1491 0.1436 0.1468 0.1451 SM(20:1) 0.0160 0.0158 0.0137 0.0141 0.0125 0.0119 0.0111 0.0131 0.0118 0.0122 0.0136 0.0125 0.0112 0.0105 0.0121 0.0122 0.0103 0.0118 0.0116 0.0138 0.0126 0.0114 0.0121 0.0112 SM(22:0) 0.4780 0.4501 0.4398 0.4198 0.4576 0.4140 0.4188 0.4743 0.5094 0.5222 0.5923 0.5368 0.4424 0.4324 0.4631 0.4797 0.3621 0.3979 0.3890 0.4442 0.3801 0.3678 0.3844 0.3722 SM(22:1) 0.0955 0.0895 0.0834 0.0823 0.0715 0.0714 0.0671 0.0803 0.0788 0.0810 0.0890 0.0830 0.0712 0.0678 0.0756 0.0811 0.0624 0.0686 0.0668 0.0772 0.0688 0.0627 0.0710 0.0666 SM(24:0) 0.2645 0.2584 0.2569 0.2294 0.2106 0.1848 0.2121 0.2163 0.2614 0.2628 0.3100 0.2627 0.2166 0.2206 0.2190 0.2212 0.1708 0.1854 0.1837 0.2072 0.1676 0.1730 0.1956 0.1776 SM(24:1) 0.3702 0.3613 0.3416 0.3261 0.2209 0.2133 0.2299 0.2513 0.2735 0.2647 0.3002 0.2792 0.2400 0.2448 0.2456 0.2675 0.2056 0.2231 0.2225 0.2611 0.2105 0.2120 0.2415 0.2206 SM(26:0) 0.0175 0.0165 0.0163 0.0143 0.0130 0.0115 0.0126 0.0133 0.0181 0.0189 0.0230 0.0190 0.0138 0.0132 0.0137 0.0141 0.0112 0.0119 0.0120 0.0125 0.0101 0.0107 0.0112 0.0107 SM(26:1) 0.0238 0.0228 0.0219 0.0203 0.0136 0.0129 0.0136 0.0156 0.0177 0.0171 0.0203 0.0186 0.0154 0.0142 0.0157 0.0162 0.0116 0.0136 0.0131 0.0140 0.0126 0.0123 0.0135 0.0132 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 SM(14:0) SM(16:0) SM(18:0) SM(18:1) SM(20:0) SM(20:1) SM(22:0) SM(22:1) SM(24:0) SM(24:1) SM(26:0) SM(26:1) METABOLITES_END #END