#METABOLOMICS WORKBENCH Kugan91_20250529_183719 DATATRACK_ID:5970 STUDY_ID:ST003962 ANALYSIS_ID:AN006510 PROJECT_ID:PR002482
VERSION             	1
CREATED_ON             	June 1, 2025, 6:05 pm
#PROJECT
PR:PROJECT_TITLE                 	Complex II assembly drives metabolic adaptation to OXPHOS dysfunction.
PR:PROJECT_SUMMARY               	During acute oxidative phosphorylation (OXPHOS) dysfunction, the reverse
PR:PROJECT_SUMMARY               	activity of succinate dehydrogenase (Complex II) maintains the redox state of
PR:PROJECT_SUMMARY               	Coenzyme Q by utilizing either fumarate or oxygen as terminal electron
PR:PROJECT_SUMMARY               	acceptors. The tendency for one over another has been suggested to be
PR:PROJECT_SUMMARY               	tissue-specific, but the underlying mechanism and consequence of this is
PR:PROJECT_SUMMARY               	unknown. Using quantitative proteomics to screen a panel of HEK293T knockout
PR:PROJECT_SUMMARY               	cell lines, we identified an increase in SDHAF2 protein, a Complex II assembly
PR:PROJECT_SUMMARY               	factor that enhances the flavination of catalytic subunit SDHA, as critical for
PR:PROJECT_SUMMARY               	metabolic adaptation during OXPHOS stress in HEK293T cells. Loss of SDHAF2
PR:PROJECT_SUMMARY               	during Complex III inhibition resulted in a reduction in Complex II F-site
PR:PROJECT_SUMMARY               	derived reactive oxygen species (ROS), a severe growth impairment, and a net
PR:PROJECT_SUMMARY               	reductive TCA cycle driven by an inability of mitochondria to support additional
PR:PROJECT_SUMMARY               	Complex II assembly. This in turn leads to use of fumarate as terminal electron
PR:PROJECT_SUMMARY               	acceptor at the cost of a ROS-mediated switch to glycolysis. Cell lines adapted
PR:PROJECT_SUMMARY               	to glycolysis did not accumulate SDHAF2 upon OXPHOS stress and exhibited a net
PR:PROJECT_SUMMARY               	reductive TCA cycle and mild growth phenotypes with or without SDHAF2 being
PR:PROJECT_SUMMARY               	present. Thus, our study reveals how Complex II assembly controls a balance
PR:PROJECT_SUMMARY               	between protection of the Q-pool and ROS-meditated signaling during oxidative
PR:PROJECT_SUMMARY               	stress in cells reliant on mitochondrial OXPHOS.
PR:INSTITUTE                     	University of Melbourne
PR:LAST_NAME                     	Roopasingam
PR:FIRST_NAME                    	Kugapreethan
PR:ADDRESS                       	30 Flemington Rd, Bio21 Molecular Science and Biotechnology Institute,
PR:ADDRESS                       	University of Melbourne, Parkville, VIC, Australia.
PR:EMAIL                         	k.roopasingam@unimelb.edu.au
PR:PHONE                         	0434297212
#STUDY
ST:STUDY_TITLE                   	Measuring TCA cycle directional fluxes using 13C5-glutamine
ST:STUDY_SUMMARY                 	This study investigates the directionality of TCA cycle flux at Complex II in
ST:STUDY_SUMMARY                 	various cell lines exhibiting OXPHOS dysfunction using targeted GS-MS analysis.
ST:INSTITUTE                     	University of Melbourne
ST:LAST_NAME                     	Roopasingam
ST:FIRST_NAME                    	Kugapreethan
ST:ADDRESS                       	30 Flemington Rd, Bio21 Molecular Science and Biotechnology Institute,
ST:ADDRESS                       	University of Melbourne, Parkville, VIC, Australia.
ST:EMAIL                         	k.roopasingam@unimelb.edu.au
ST:PHONE                         	0434297212
#SUBJECT
SU:SUBJECT_TYPE                  	Cultured cells
SU:SUBJECT_SPECIES               	Homo sapiens
SU:TAXONOMY_ID                   	9606
#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           	-	03_HEKWT_MRM_001_UNK-0011_16042025_11	Sample source:HEK_WT_001 | Treatment:HEK_WT	RAW_FILE_NAME(Raw file name)=03_HEKWT_MRM_001_UNK-0011_16042025_11.qgd
SUBJECT_SAMPLE_FACTORS           	-	03_HEKWT_MRM_002_UNK-0009_16042025_9	Sample source:HEK_WT_002 | Treatment:HEK_WT	RAW_FILE_NAME(Raw file name)=03_HEKWT_MRM_002_UNK-0009_16042025_9.qgd
SUBJECT_SAMPLE_FACTORS           	-	03_HEKWT_MRM_003_UNK-0010_16042025_10	Sample source:HEK_WT_003 | Treatment:HEK_WT	RAW_FILE_NAME(Raw file name)=03_HEKWT_MRM_003_UNK-0010_16042025_10.qgd
SUBJECT_SAMPLE_FACTORS           	-	04_HEK_A-A_MRM_001_UNK-0014_16042025_14	Sample source:HEK_A-A_001 | Treatment:HEK_A-A	RAW_FILE_NAME(Raw file name)=04_HEK_A-A_MRM_001_UNK-0014_16042025_14.qgd
SUBJECT_SAMPLE_FACTORS           	-	04_HEK_A-A_MRM_002_UNK-0012_16042025_12	Sample source:HEK_A-A_002 | Treatment:HEK_A-A	RAW_FILE_NAME(Raw file name)=04_HEK_A-A_MRM_002_UNK-0012_16042025_12.qgd
SUBJECT_SAMPLE_FACTORS           	-	04_HEK_A-A_MRM_003_UNK-0013_16042025_13	Sample source:HEK_A-A_003 | Treatment:HEK_A-A	RAW_FILE_NAME(Raw file name)=04_HEK_A-A_MRM_003_UNK-0013_16042025_13.qgd
SUBJECT_SAMPLE_FACTORS           	-	05_HEKSDHAF2_MRM_001_UNK-0017_16042025_17	Sample source:HEKSDHAF2KO_001 | Treatment:HEKSDHAF2KO	RAW_FILE_NAME(Raw file name)=05_HEKSDHAF2_MRM_001_UNK-0017_16042025_17.qgd
SUBJECT_SAMPLE_FACTORS           	-	05_HEKSDHAF2_MRM_002_UNK-0015_16042025_15	Sample source:HEKSDHAF2KO_002 | Treatment:HEKSDHAF2KO	RAW_FILE_NAME(Raw file name)=05_HEKSDHAF2_MRM_002_UNK-0015_16042025_15.qgd
SUBJECT_SAMPLE_FACTORS           	-	05_HEKSDHAF2_MRM_003_UNK-0016_16042025_16	Sample source:HEKSDHAF2KO_003 | Treatment:HEKSDHAF2KO	RAW_FILE_NAME(Raw file name)=05_HEKSDHAF2_MRM_003_UNK-0016_16042025_16.qgd
SUBJECT_SAMPLE_FACTORS           	-	06_HEKSDHAF2_A-A_MRM_001_SET1_UNK-0020_20042025_6	Sample source:HEKSDHAF2KO_A-A_001 | Treatment:HEKSDHAF2KO_A-A	RAW_FILE_NAME(Raw file name)=06_HEKSDHAF2_A-A_MRM_001_SET1_UNK-0020_20042025_6.qgd
SUBJECT_SAMPLE_FACTORS           	-	06_HEKSDHAF2_A-A_MRM_001_SET2_UNK-0027_20042025_9	Sample source:HEKSDHAF2KO_A-A_002 | Treatment:HEKSDHAF2KO_A-A	RAW_FILE_NAME(Raw file name)=06_HEKSDHAF2_A-A_MRM_001_SET2_UNK-0027_20042025_9.qgd
SUBJECT_SAMPLE_FACTORS           	-	06_HEKSDHAF2_A-A_MRM_003_SET2_UNK-0026_20042025_8	Sample source:HEKSDHAF2KO_A-A_003 | Treatment:HEKSDHAF2KO_A-A	RAW_FILE_NAME(Raw file name)=06_HEKSDHAF2_A-A_MRM_003_SET2_UNK-0026_20042025_8.qgd
SUBJECT_SAMPLE_FACTORS           	-	07_SDHAF2_SDHAF2FLAG_MRM_001_UNK-0023_16042025_23	Sample source:HEKSDHAF2_SDHAF2FLAG_001 | Treatment:HEKSDHAF2_SDHAF2FLAG	RAW_FILE_NAME(Raw file name)=07_SDHAF2_SDHAF2FLAG_MRM_001_UNK-0023_16042025_23.qgd
SUBJECT_SAMPLE_FACTORS           	-	07_SDHAF2_SDHAF2FLAG_MRM_002_UNK-0021_16042025_21	Sample source:HEKSDHAF2_SDHAF2FLAG_002 | Treatment:HEKSDHAF2_SDHAF2FLAG	RAW_FILE_NAME(Raw file name)=07_SDHAF2_SDHAF2FLAG_MRM_002_UNK-0021_16042025_21.qgd
SUBJECT_SAMPLE_FACTORS           	-	07_SDHAF2_SDHAF2FLAG_MRM_003_UNK-0022_16042025_22	Sample source:HEKSDHAF2_SDHAF2FLAG_003 | Treatment:HEKSDHAF2_SDHAF2FLAG	RAW_FILE_NAME(Raw file name)=07_SDHAF2_SDHAF2FLAG_MRM_003_UNK-0022_16042025_22.qgd
SUBJECT_SAMPLE_FACTORS           	-	08_SDHAF2_CIIIKO_A-A_MRM_001_RERUN_SET2_UNK-0030_20042025_15	Sample source:HEKSDHAF2KO_CIIIKO_A-A_001 | Treatment:HEKSDHAF2KO_CIIIKO_A-A	RAW_FILE_NAME(Raw file name)=08_SDHAF2_CIIIKO_A-A_MRM_001_RERUN_SET2_UNK-0030_20042025_15.qgd
SUBJECT_SAMPLE_FACTORS           	-	08_SDHAF2_CIIIKO_A-A_MRM_001_UNK-0027_16042025_27	Sample source:HEKSDHAF2KO_CIIIKO_A-A_002 | Treatment:HEKSDHAF2KO_CIIIKO_A-A	RAW_FILE_NAME(Raw file name)=08_SDHAF2_CIIIKO_A-A_MRM_001_UNK-0027_16042025_27.qgd
SUBJECT_SAMPLE_FACTORS           	-	08_SDHAF2_CIIIKO_A-A_MRM_003_RERUN_SET2_UNK-0029_20042025_14	Sample source:HEKSDHAF2KO_CIIIKO_A-A_003 | Treatment:HEKSDHAF2KO_CIIIKO_A-A	RAW_FILE_NAME(Raw file name)=08_SDHAF2_CIIIKO_A-A_MRM_003_RERUN_SET2_UNK-0029_20042025_14.qgd
SUBJECT_SAMPLE_FACTORS           	-	13_HAP_MRM_001_UNK-0042_16042025_42	Sample source:HAP_WT_001 | Treatment:HAP_WT	RAW_FILE_NAME(Raw file name)=13_HAP_MRM_001_UNK-0042_16042025_42.qgd
SUBJECT_SAMPLE_FACTORS           	-	13_HAP_MRM_002_UNK-0040_16042025_40	Sample source:HAP_WT_002 | Treatment:HAP_WT	RAW_FILE_NAME(Raw file name)=13_HAP_MRM_002_UNK-0040_16042025_40.qgd
SUBJECT_SAMPLE_FACTORS           	-	13_HAP_MRM_003_UNK-0041_16042025_41	Sample source:HAP_WT_003 | Treatment:HAP_WT	RAW_FILE_NAME(Raw file name)=13_HAP_MRM_003_UNK-0041_16042025_41.qgd
SUBJECT_SAMPLE_FACTORS           	-	14_HAP_A-A_MRM_001_UNK-0046_16042025_46	Sample source:HAP_A-A_001 | Treatment:HAP_A-A	RAW_FILE_NAME(Raw file name)=14_HAP_A-A_MRM_001_UNK-0046_16042025_46.qgd
SUBJECT_SAMPLE_FACTORS           	-	14_HAP_A-A_MRM_002_UNK-0044_16042025_44	Sample source:HAP_A-A_002 | Treatment:HAP_A-A	RAW_FILE_NAME(Raw file name)=14_HAP_A-A_MRM_002_UNK-0044_16042025_44.qgd
SUBJECT_SAMPLE_FACTORS           	-	14_HAP_A-A_MRM_003_UNK-0045_16042025_45	Sample source:HAP_A-A_003 | Treatment:HAP_A-A	RAW_FILE_NAME(Raw file name)=14_HAP_A-A_MRM_003_UNK-0045_16042025_45.qgd
SUBJECT_SAMPLE_FACTORS           	-	19_HeLa_MRM_001_UNK-0058_16042025_58	Sample source:HeLa_WT_001 | Treatment:HeLa_WT	RAW_FILE_NAME(Raw file name)=19_HeLa_MRM_001_UNK-0058_16042025_58.qgd
SUBJECT_SAMPLE_FACTORS           	-	19_HeLa_MRM_002_UNK-0056_16042025_56	Sample source:HeLa_WT_002 | Treatment:HeLa_WT	RAW_FILE_NAME(Raw file name)=19_HeLa_MRM_002_UNK-0056_16042025_56.qgd
SUBJECT_SAMPLE_FACTORS           	-	19_HeLa_MRM_003_UNK-0057_16042025_57	Sample source:HeLa_WT_003 | Treatment:HeLa_WT	RAW_FILE_NAME(Raw file name)=19_HeLa_MRM_003_UNK-0057_16042025_57.qgd
SUBJECT_SAMPLE_FACTORS           	-	20_HeLa_A-A_MRM_001_UNK-0061_16042025_61	Sample source:HeLa_A-A_001 | Treatment:HeLa_A-A	RAW_FILE_NAME(Raw file name)=20_HeLa_A-A_MRM_001_UNK-0061_16042025_61.qgd
SUBJECT_SAMPLE_FACTORS           	-	20_HeLa_A-A_MRM_002_UNK-0059_16042025_59	Sample source:HeLa_A-A_002 | Treatment:HeLa_A-A	RAW_FILE_NAME(Raw file name)=20_HeLa_A-A_MRM_002_UNK-0059_16042025_59.qgd
SUBJECT_SAMPLE_FACTORS           	-	20_HeLa_A-A_MRM_003_UNK-0060_16042025_60	Sample source:HeLa_A-A_003 | Treatment:HeLa_A-A	RAW_FILE_NAME(Raw file name)=20_HeLa_A-A_MRM_003_UNK-0060_16042025_60.qgd
SUBJECT_SAMPLE_FACTORS           	-	25_MCF7_MRM_001_UNK-0074_16042025_74	Sample source:MCF7_WT_001 | Treatment:MCF7_WT	RAW_FILE_NAME(Raw file name)=25_MCF7_MRM_001_UNK-0074_16042025_74.qgd
SUBJECT_SAMPLE_FACTORS           	-	25_MCF7_MRM_002_UNK-0072_16042025_72	Sample source:MCF7_WT_002 | Treatment:MCF7_WT	RAW_FILE_NAME(Raw file name)=25_MCF7_MRM_002_UNK-0072_16042025_72.qgd
SUBJECT_SAMPLE_FACTORS           	-	25_MCF7_MRM_003_UNK-0073_16042025_73	Sample source:MCF7_WT_003 | Treatment:MCF7_WT	RAW_FILE_NAME(Raw file name)=25_MCF7_MRM_003_UNK-0073_16042025_73.qgd
SUBJECT_SAMPLE_FACTORS           	-	26_MCF7_A-A_MRM_001_UNK-0077_16042025_77	Sample source:MCF7_A-A_001 | Treatment:MCF7_A-A	RAW_FILE_NAME(Raw file name)=26_MCF7_A-A_MRM_001_UNK-0077_16042025_77.qgd
SUBJECT_SAMPLE_FACTORS           	-	26_MCF7_A-A_MRM_002_UNK-0075_16042025_75	Sample source:MCF7_A-A_002 | Treatment:MCF7_A-A	RAW_FILE_NAME(Raw file name)=26_MCF7_A-A_MRM_002_UNK-0075_16042025_75.qgd
SUBJECT_SAMPLE_FACTORS           	-	26_MCF7_A-A_MRM_003_UNK-0076_16042025_76	Sample source:MCF7_A-A_003 | Treatment:MCF7_A-A	RAW_FILE_NAME(Raw file name)=26_MCF7_A-A_MRM_003_UNK-0076_16042025_76.qgd
SUBJECT_SAMPLE_FACTORS           	-	27_143B_WT_MRM_028_UNK-0018_9042025_18	Sample source:143B_WT_001 | Treatment:143B_WT	RAW_FILE_NAME(Raw file name)=27_143B_WT_MRM_028_UNK-0018_9042025_18.qgd
SUBJECT_SAMPLE_FACTORS           	-	27_143B_WT_MRM_030_UNK-0020_9042025_20	Sample source:143B_WT_002 | Treatment:143B_WT	RAW_FILE_NAME(Raw file name)=27_143B_WT_MRM_030_UNK-0020_9042025_20.qgd
SUBJECT_SAMPLE_FACTORS           	-	27_143B_WT_MRM__029_UNK-0019_9042025_19	Sample source:143B_WT_003 | Treatment:143B_WT	RAW_FILE_NAME(Raw file name)=27_143B_WT_MRM__029_UNK-0019_9042025_19.qgd
SUBJECT_SAMPLE_FACTORS           	-	28_143B_A-A_MRM_034_UNK-0021_9042025_21	Sample source:143B_A_A_001 | Treatment:143B_A_A	RAW_FILE_NAME(Raw file name)=28_143B_A-A_MRM_034_UNK-0021_9042025_21.qgd
SUBJECT_SAMPLE_FACTORS           	-	28_143B_A-A_MRM__035_UNK-0022_9042025_22	Sample source:143B_A_A_002 | Treatment:143B_A_A	RAW_FILE_NAME(Raw file name)=28_143B_A-A_MRM__035_UNK-0022_9042025_22.qgd
SUBJECT_SAMPLE_FACTORS           	-	28_143B_A-A_MRM__036_UNK-0023_9042025_23	Sample source:143B_A_A_003 | Treatment:143B_A_A	RAW_FILE_NAME(Raw file name)=28_143B_A-A_MRM__036_UNK-0023_9042025_23.qgd
#COLLECTION
CO:COLLECTION_SUMMARY            	HEK293T. HeLa, eHAP, MCF7 and 143B cells were cultured in either Dulbecco's
CO:COLLECTION_SUMMARY            	Modified Eagle Medium (DMEM, Thermo Fisher Scientific) or IMDM media,
CO:COLLECTION_SUMMARY            	supplemented with 10 % (v/v) fetal bovine serum (FBS, CellSera), 50 µg/mL
CO:COLLECTION_SUMMARY            	uridine (Sigma), and a mixture of 100 µg/mL streptomycin and 100 units/mL
CO:COLLECTION_SUMMARY            	penicillin (Thermo Scientific). For stable isotopologue tracing of
CO:COLLECTION_SUMMARY            	13C5-glutamine (Merck), media was replaced with glutamine-free DMEM (Thermo
CO:COLLECTION_SUMMARY            	fisher Scientific) or IMDM supplemented with 2 mM of 13C5-glutamine, 1 mM
CO:COLLECTION_SUMMARY            	pyruvate (Thermo Scientific), 10% FBS, 50 µg/mL uridine, and a mixture of 100
CO:COLLECTION_SUMMARY            	µg/mL streptomycin and 100 units/mL penicillin.
CO:SAMPLE_TYPE                   	Cultured cells
#TREATMENT
TR:TREATMENT_SUMMARY             	Relevant cell lines were treated with doxycycline and Antimycin A 24 and 8 hours
TR:TREATMENT_SUMMARY             	prior to the addition of tracer media and maintained until the completion of the
TR:TREATMENT_SUMMARY             	experiment, respectively.
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	Polar metabolites were extracted from snap-frozen cells using 600 µL of HPLC
SP:SAMPLEPREP_SUMMARY            	grade methanol:chloroform mixture (9:1; [v/v]), along with internal standards
SP:SAMPLEPREP_SUMMARY            	(1.66μM 13C5,15N-valine, 1.66μM 13C6-sorbitol), as described above. The
SP:SAMPLEPREP_SUMMARY            	clarified supernatants and pooled biological quality controls (PBQC’s) were
SP:SAMPLEPREP_SUMMARY            	dried using a CentriVap concentrator (Labconco).
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_TYPE           	GC
CH:INSTRUMENT_NAME               	Shimadzu GC-2010
CH:COLUMN_NAME                   	30 m Agilent DB-5 column with a diameter of 0.25mm and a film thickness of 1µm
CH:SOLVENT_A                     	N/A
CH:SOLVENT_B                     	N/A
CH:FLOW_GRADIENT                 	N/A
CH:FLOW_RATE                     	1ml/min
CH:COLUMN_TEMPERATURE            	100
#ANALYSIS
AN:ANALYSIS_TYPE                 	MS
#MS
MS:INSTRUMENT_NAME               	Shimadzu TQ8050NX
MS:INSTRUMENT_TYPE               	Triple quadrupole
MS:MS_TYPE                       	EI
MS:ION_MODE                      	POSITIVE
MS:MS_COMMENTS                   	The GC-MS system used comprised of an AOC6000 autosampler, a 2030 Shimadzu gas
MS:MS_COMMENTS                   	chromatograph and a TQ8050NX triple quadrupole mass spectrometer (Shimadzu,
MS:MS_COMMENTS                   	Japan)with an electron ionisation source(-70eV). The mass spectrometer was tuned
MS:MS_COMMENTS                   	according to the manufacturer’s recommendations using
MS:MS_COMMENTS                   	tris-(perfluorobutyl)-amine (CF43). GC-MS was performed on a 30m Agilent DB-5
MS:MS_COMMENTS                   	column with 0.25mm internal diameter column and 1µm film thickness. The
MS:MS_COMMENTS                   	injection temperature (inlet) was set at 280°C, the MS transfer line at 280°C
MS:MS_COMMENTS                   	and the ion source adjusted to 200°C. Helium was used as the carrier gas at a
MS:MS_COMMENTS                   	flow rate of 1 mL/min and argon gas was used in the collision cell to generate
MS:MS_COMMENTS                   	the MRM product ion. The analysis of the derivatised samples was performed under
MS:MS_COMMENTS                   	the following oven temperature program; 100°C start temperature, hold for 4
MS:MS_COMMENTS                   	minutes, followed by a 10°C/min oven temperature ramp to 320°C with a
MS:MS_COMMENTS                   	following final hold for 11 minutes.
#MS_METABOLITE_DATA
MS_METABOLITE_DATA:UNITS	Relative abundance
MS_METABOLITE_DATA_START
Samples	03_HEKWT_MRM_001_UNK-0011_16042025_11	03_HEKWT_MRM_002_UNK-0009_16042025_9	03_HEKWT_MRM_003_UNK-0010_16042025_10	04_HEK_A-A_MRM_001_UNK-0014_16042025_14	04_HEK_A-A_MRM_002_UNK-0012_16042025_12	04_HEK_A-A_MRM_003_UNK-0013_16042025_13	05_HEKSDHAF2_MRM_001_UNK-0017_16042025_17	05_HEKSDHAF2_MRM_002_UNK-0015_16042025_15	05_HEKSDHAF2_MRM_003_UNK-0016_16042025_16	06_HEKSDHAF2_A-A_MRM_001_SET1_UNK-0020_20042025_6	06_HEKSDHAF2_A-A_MRM_001_SET2_UNK-0027_20042025_9	06_HEKSDHAF2_A-A_MRM_003_SET2_UNK-0026_20042025_8	07_SDHAF2_SDHAF2FLAG_MRM_001_UNK-0023_16042025_23	07_SDHAF2_SDHAF2FLAG_MRM_002_UNK-0021_16042025_21	07_SDHAF2_SDHAF2FLAG_MRM_003_UNK-0022_16042025_22	08_SDHAF2_CIIIKO_A-A_MRM_001_RERUN_SET2_UNK-0030_20042025_15	08_SDHAF2_CIIIKO_A-A_MRM_001_UNK-0027_16042025_27	08_SDHAF2_CIIIKO_A-A_MRM_003_RERUN_SET2_UNK-0029_20042025_14	13_HAP_MRM_001_UNK-0042_16042025_42	13_HAP_MRM_002_UNK-0040_16042025_40	13_HAP_MRM_003_UNK-0041_16042025_41	14_HAP_A-A_MRM_001_UNK-0046_16042025_46	14_HAP_A-A_MRM_002_UNK-0044_16042025_44	14_HAP_A-A_MRM_003_UNK-0045_16042025_45	19_HeLa_MRM_001_UNK-0058_16042025_58	19_HeLa_MRM_002_UNK-0056_16042025_56	19_HeLa_MRM_003_UNK-0057_16042025_57	20_HeLa_A-A_MRM_001_UNK-0061_16042025_61	20_HeLa_A-A_MRM_002_UNK-0059_16042025_59	20_HeLa_A-A_MRM_003_UNK-0060_16042025_60	25_MCF7_MRM_001_UNK-0074_16042025_74	25_MCF7_MRM_002_UNK-0072_16042025_72	25_MCF7_MRM_003_UNK-0073_16042025_73	26_MCF7_A-A_MRM_001_UNK-0077_16042025_77	26_MCF7_A-A_MRM_002_UNK-0075_16042025_75	26_MCF7_A-A_MRM_003_UNK-0076_16042025_76	27_143B_WT_MRM_028_UNK-0018_9042025_18	27_143B_WT_MRM_030_UNK-0020_9042025_20	27_143B_WT_MRM__029_UNK-0019_9042025_19	28_143B_A-A_MRM_034_UNK-0021_9042025_21	28_143B_A-A_MRM__035_UNK-0022_9042025_22	28_143B_A-A_MRM__036_UNK-0023_9042025_23
Factors	Sample source:HEK_WT_001 | Treatment:HEK_WT	Sample source:HEK_WT_002 | Treatment:HEK_WT	Sample source:HEK_WT_003 | Treatment:HEK_WT	Sample source:HEK_A-A_001 | Treatment:HEK_A-A	Sample source:HEK_A-A_002 | Treatment:HEK_A-A	Sample source:HEK_A-A_003 | Treatment:HEK_A-A	Sample source:HEKSDHAF2KO_001 | Treatment:HEKSDHAF2KO	Sample source:HEKSDHAF2KO_002 | Treatment:HEKSDHAF2KO	Sample source:HEKSDHAF2KO_003 | Treatment:HEKSDHAF2KO	Sample source:HEKSDHAF2KO_A-A_001 | Treatment:HEKSDHAF2KO_A-A	Sample source:HEKSDHAF2KO_A-A_002 | Treatment:HEKSDHAF2KO_A-A	Sample source:HEKSDHAF2KO_A-A_003 | Treatment:HEKSDHAF2KO_A-A	Sample source:HEKSDHAF2_SDHAF2FLAG_001 | Treatment:HEKSDHAF2_SDHAF2FLAG	Sample source:HEKSDHAF2_SDHAF2FLAG_002 | Treatment:HEKSDHAF2_SDHAF2FLAG	Sample source:HEKSDHAF2_SDHAF2FLAG_003 | Treatment:HEKSDHAF2_SDHAF2FLAG	Sample source:HEKSDHAF2KO_CIIIKO_A-A_001 | Treatment:HEKSDHAF2KO_CIIIKO_A-A	Sample source:HEKSDHAF2KO_CIIIKO_A-A_002 | Treatment:HEKSDHAF2KO_CIIIKO_A-A	Sample source:HEKSDHAF2KO_CIIIKO_A-A_003 | Treatment:HEKSDHAF2KO_CIIIKO_A-A	Sample source:HAP_WT_001 | Treatment:HAP_WT	Sample source:HAP_WT_002 | Treatment:HAP_WT	Sample source:HAP_WT_003 | Treatment:HAP_WT	Sample source:HAP_A-A_001 | Treatment:HAP_A-A	Sample source:HAP_A-A_002 | Treatment:HAP_A-A	Sample source:HAP_A-A_003 | Treatment:HAP_A-A	Sample source:HeLa_WT_001 | Treatment:HeLa_WT	Sample source:HeLa_WT_002 | Treatment:HeLa_WT	Sample source:HeLa_WT_003 | Treatment:HeLa_WT	Sample source:HeLa_A-A_001 | Treatment:HeLa_A-A	Sample source:HeLa_A-A_002 | Treatment:HeLa_A-A	Sample source:HeLa_A-A_003 | Treatment:HeLa_A-A	Sample source:MCF7_WT_001 | Treatment:MCF7_WT	Sample source:MCF7_WT_002 | Treatment:MCF7_WT	Sample source:MCF7_WT_003 | Treatment:MCF7_WT	Sample source:MCF7_A-A_001 | Treatment:MCF7_A-A	Sample source:MCF7_A-A_002 | Treatment:MCF7_A-A	Sample source:MCF7_A-A_003 | Treatment:MCF7_A-A	Sample source:143B_WT_001 | Treatment:143B_WT	Sample source:143B_WT_002 | Treatment:143B_WT	Sample source:143B_WT_003 | Treatment:143B_WT	Sample source:143B_A_A_001 | Treatment:143B_A_A	Sample source:143B_A_A_002 | Treatment:143B_A_A	Sample source:143B_A_A_003 | Treatment:143B_A_A
Succinic acid-2TMS_247.1> 147.1	2454070.7	2545901.4	2438531.2	11178418.8	10996823.4	10518073.3	3276572.2	10772778.5	3427849.8	1401638.3	1293987.4	3383607.2	2426138.7	2526766.7	2237251.7	3397848.4	2057345.3	5643343.8	1489506.2	1456624.4	1482331.6	11290437.4	15552077.8	14768773.1	1676909.9	2323084.3	2648756.7	7365082	9795108.6	10341682.5	3290519.8	3927078.2	3899777	10130661.2	10697995.7	10384937.8	26079.2	28617.5	34471.2	197566.8	208042.8	205104
Succinic acid-2TMS_248.1> 147.1	396688.9	394148.9	359301.5	339534.8	300916.2	266670.5	627445.7	2205663.8	642488.4	53580.2	54827.3	97893.8	388800.6	448472.5	357323.7	126577.4	65590.9	167953.1	210273.6	200945.3	206533.5	331931.7	453047.4	416406.2	192739.6	264080.6	280485.9	239452.6	285675.5	278572.7	682500.4	793982.8	733645.9	314154.5	316156.2	341536.8	7036.2	7403.6	9030.9	7202.3	7142.4	7234.5
Succinic acid-2TMS_249.1> 147.1	755117.4	714043	665731.7	-157365.9	-141530.7	-118615.6	1023733.9	3634239.4	1065668.3	-1820.5	843.1	-9776.7	726894.9	768297.7	620462.9	-38697.3	-33539.4	-76909.5	407977.6	437416.8	414544.7	-170251.9	-207155.8	-193991	776805	1001247.3	1079569	-131713.6	-159749.9	-176307.8	704541.3	884049.4	802156.5	558418.7	578212.5	566947.7	15899.3	17138.2	21556.1	17915.1	19164.8	19308.4
Succinic acid-2TMS_250.1> 147.1	64329.6	64275.6	58762.3	2629279.9	2715027.2	2480296.6	101504	317032	99912.1	217308.1	198523.5	517396.2	58408.4	71955.8	57582.6	554036.3	333391	930443.8	52294.3	53141.9	48568.8	3677286.2	5268453.2	5052105.7	136910.4	186319.1	194024	1393506.4	1939945.2	2018146.1	24175.5	39577.1	48622.8	4257365.7	4775247.1	4484599.1	1304.9	1531.3	1900.4	257378.8	260075.6	258625.4
Succinic acid-2TMS_251.1> 147.1	1864911.1	1924190	1742294.2	23642114.4	23409603.4	22191919.7	2947253.1	10708610.4	3100366.3	4557548.3	4237791.4	11287487.8	1984507.4	2229913.4	1869908.4	8688150	4978047.3	14205219.7	1342667.8	1401148.6	1309055.6	11500246.9	15739320.8	14771651.6	3953115.4	5418464	5941146.1	21150468.7	24244169.7	24558240.4	1305837.4	1618968.2	1479878.7	17221974.1	18167898.8	17584319.2	42053.1	46551.8	58243.2	577379.3	631941	625461.3
Fumaric acid-2TMS_245.0> 147.1	3381128.4	3734166	2966290.7	716511	864859.6	799461.1	1109600.1	3963760.6	1122873.5	75370	141956.6	199703.7	1274142.3	1444727	1301539.6	252919	128863.2	246234.5	2515302.5	2659924.2	3249639.9	5100464	6980963.8	7002163.1	1362252.4	2019395.7	2304681.8	1350480.6	1965705.2	1796830.4	1102319.6	1571358	1286402.4	2492409.4	2793430.2	2520355.3	77169.2	81462.3	84935.1	62847	62399.7	59108.8
Fumaric acid-2TMS_246.0> 147.1	540447.6	601442.1	471155.3	29593	32625.5	35146.6	172734.2	590415.8	176625.6	1718.8	6086.4	7756.2	220611.9	237641.7	210077.9	12282.2	3804	10013.3	352104.7	393284.6	450498.1	127926.8	175555.8	169848	148809	197804.5	208846.8	42733.2	63680.2	54492.2	225483.4	317322.2	240913.1	79671.6	88373.6	76332.1	15663.1	16119.7	17136.2	2684.9	2308.3	2103.6
Fumaric acid-2TMS_247.0> 147.1	798671.5	866633.9	670395.3	-13470.3	-11726.4	-14485.5	228987.6	787441.6	227474	842.8	-3824.5	-3908.3	307920	353887.4	271510.7	-4600.3	6914	317.6	572292.3	674011.6	752291.1	-75730.2	-95604.5	-94444.7	348347.3	489037.9	457373.5	-6740.7	-7090.7	-2392.1	184103.4	256675.7	206918.9	83671.2	98299.1	88683.9	24429	25433.8	26671.5	4501.4	4386.3	4130.9
Fumaric acid-2TMS_248.0> 147.1	244322.5	272713.2	214324.1	341363.7	419783.8	356096.8	63940.7	248569.7	63554.4	15738.9	30609.7	45660.5	115082	118365.7	100696.1	78506.4	57987.7	71151.3	460602.5	518099.2	598351.1	2460276.4	3470068.4	3530984.5	333044	504451.7	510949.8	763927.4	1138362.1	1077641	106298.7	182879.8	137142.9	2353394.8	2765749.6	2414503.5	10983.2	12095.9	12346.2	111898.5	106243.7	99776.2
Fumaric acid-2TMS_249.0> 147.1	2536251.1	2836691.8	2168483.8	245986	301179.7	283858.9	721906.3	2530910.2	725203.5	6916.3	16767.7	22915.6	975641.2	1081417.9	904619.2	51038.4	35445.9	50877.1	1995267.2	2280700.8	2626881.8	639887.7	866032.6	862668.4	2143650.8	3165599.4	3216682.8	165359.9	241416.6	224273.1	378972.2	533063.7	402957.8	411964.2	482615.8	441976.9	79556	83876.8	87204.7	34995.6	33220.2	31130.7
MS_METABOLITE_DATA_END
#METABOLITES
METABOLITES_START
metabolite_name	HMDB ID
Succinic acid-2TMS_247.1> 147.1	HMDB0000254
Succinic acid-2TMS_248.1> 147.1	HMDB0000254
Succinic acid-2TMS_249.1> 147.1	HMDB0000254
Succinic acid-2TMS_250.1> 147.1	HMDB0000254
Succinic acid-2TMS_251.1> 147.1	HMDB0000254
Fumaric acid-2TMS_245.0> 147.1	HMDB0000134
Fumaric acid-2TMS_246.0> 147.1	HMDB0000134
Fumaric acid-2TMS_247.0> 147.1	HMDB0000134
Fumaric acid-2TMS_248.0> 147.1	HMDB0000134
Fumaric acid-2TMS_249.0> 147.1	HMDB0000134
METABOLITES_END
#END