#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 manufacturer’s 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            	manufacturer’s 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