#METABOLOMICS WORKBENCH yuta624_20250430_173919 DATATRACK_ID:5874 STUDY_ID:ST003905 ANALYSIS_ID:AN006410 PROJECT_ID:PR002444
VERSION             	1
CREATED_ON             	May 7, 2025, 10:14 pm
#PROJECT
PR:PROJECT_TITLE                 	Lysosomal dysfunction impairs lipid peroxidation and ferroptosis in senescent
PR:PROJECT_TITLE                 	cells
PR:PROJECT_SUMMARY               	Senescent cells, characterized by irreversible cell cycle arrest and the
PR:PROJECT_SUMMARY               	secretion of inflammatory factors, promote various age-related pathologies1.
PR:PROJECT_SUMMARY               	Recently, multiple therapeutic approaches have been developed to eliminate
PR:PROJECT_SUMMARY               	senescent cells by targeting their cell death resistance pathways. Senescent
PR:PROJECT_SUMMARY               	cells exhibit resistance to ferroptosis, a form of iron-dependent cell death2,3;
PR:PROJECT_SUMMARY               	however, the underlying mechanisms remain unclear. Here we discovered that
PR:PROJECT_SUMMARY               	lysosomal acidity is crucial for lipid peroxidation and the induction of
PR:PROJECT_SUMMARY               	ferroptosis by cystine deprivation. In senescent cells, the expression of
PR:PROJECT_SUMMARY               	ATP6V1C2, a component of V-ATPase, is downregulated, leading to lysosomal
PR:PROJECT_SUMMARY               	dysfunction and resistance to lipid peroxidation and ferroptosis. Inactivation
PR:PROJECT_SUMMARY               	of V-ATPase by concanamycin C or knockdown of ATP6V1C2 increases lysosomal pH,
PR:PROJECT_SUMMARY               	inhibiting lipid peroxidation and ferroptosis in proliferating cells.
PR:PROJECT_SUMMARY               	Conversely, treatment with the pharmacological agent EN6 or overexpression of
PR:PROJECT_SUMMARY               	ATP6V1C2 reduces lysosomal pH and recovers ferroptosis sensitivity in senescent
PR:PROJECT_SUMMARY               	cells and therapy-induced senescent cancer cells. Furthermore, derivation of
PR:PROJECT_SUMMARY               	ferroptosis vulnerability by EN6 prevented pancreatic cancer development in
PR:PROJECT_SUMMARY               	xenograft and KC mouse models. Our findings reveal a link among lysosomal
PR:PROJECT_SUMMARY               	dysfunction, lipid peroxidation, and the regulation of ferroptosis in cellular
PR:PROJECT_SUMMARY               	senescence, suggesting that inducing ferroptosis in senescent cells by
PR:PROJECT_SUMMARY               	modulating lysosomal pH could be a therapeutic strategy for the treatment of
PR:PROJECT_SUMMARY               	age-related diseases.
PR:INSTITUTE                     	Kyoto University
PR:DEPARTMENT                    	Graduate School of Medicine
PR:LABORATORY                    	Center for Cancer Immunotherapy and Immunobiology (CCII)
PR:LAST_NAME                     	Matsuoka
PR:FIRST_NAME                    	Yuta
PR:ADDRESS                       	53 Shogoin Kawahara-cho, Sakyo-ku
PR:EMAIL                         	matsuoka.yuta.6r@kyoto-u.ac.jp
PR:PHONE                         	0757514150
#STUDY
ST:STUDY_TITLE                   	Lysosomal dysfunction impairs lipid peroxidation and ferroptosis in senescent
ST:STUDY_TITLE                   	cells
ST:STUDY_SUMMARY                 	Senescent cells, characterized by irreversible cell cycle arrest and the
ST:STUDY_SUMMARY                 	secretion of inflammatory factors, promote various age-related pathologies1.
ST:STUDY_SUMMARY                 	Recently, multiple therapeutic approaches have been developed to eliminate
ST:STUDY_SUMMARY                 	senescent cells by targeting their cell death resistance pathways. Senescent
ST:STUDY_SUMMARY                 	cells exhibit resistance to ferroptosis, a form of iron-dependent cell death2,3;
ST:STUDY_SUMMARY                 	however, the underlying mechanisms remain unclear. Here we discovered that
ST:STUDY_SUMMARY                 	lysosomal acidity is crucial for lipid peroxidation and the induction of
ST:STUDY_SUMMARY                 	ferroptosis by cystine deprivation. In senescent cells, the expression of
ST:STUDY_SUMMARY                 	ATP6V1C2, a component of V-ATPase, is downregulated, leading to lysosomal
ST:STUDY_SUMMARY                 	dysfunction and resistance to lipid peroxidation and ferroptosis. Inactivation
ST:STUDY_SUMMARY                 	of V-ATPase by concanamycin C or knockdown of ATP6V1C2 increases lysosomal pH,
ST:STUDY_SUMMARY                 	inhibiting lipid peroxidation and ferroptosis in proliferating cells.
ST:STUDY_SUMMARY                 	Conversely, treatment with the pharmacological agent EN6 or overexpression of
ST:STUDY_SUMMARY                 	ATP6V1C2 reduces lysosomal pH and recovers ferroptosis sensitivity in senescent
ST:STUDY_SUMMARY                 	cells and therapy-induced senescent cancer cells. Furthermore, derivation of
ST:STUDY_SUMMARY                 	ferroptosis vulnerability by EN6 prevented pancreatic cancer development in
ST:STUDY_SUMMARY                 	xenograft and KC mouse models. Our findings reveal a link among lysosomal
ST:STUDY_SUMMARY                 	dysfunction, lipid peroxidation, and the regulation of ferroptosis in cellular
ST:STUDY_SUMMARY                 	senescence, suggesting that inducing ferroptosis in senescent cells by
ST:STUDY_SUMMARY                 	modulating lysosomal pH could be a therapeutic strategy for the treatment of
ST:STUDY_SUMMARY                 	age-related diseases.
ST:INSTITUTE                     	Kyoto University
ST:LAST_NAME                     	Matsuoka
ST:FIRST_NAME                    	Yuta
ST:ADDRESS                       	53 Shogoin Kawahara-cho, Sakyo-ku
ST:EMAIL                         	matsuoka.yuta.6r@kyoto-u.ac.jp
ST:PHONE                         	+81757514150
#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           	-	sample 1	Sample source:Cultured cells | Cell type:normal	Treatment=DMSO; RAW_FILE_NAME(Raw file name)=sample 1.mzML
SUBJECT_SAMPLE_FACTORS           	-	sample 2	Sample source:Cultured cells | Cell type:normal	Treatment=DMSO; RAW_FILE_NAME(Raw file name)=sample 2.mzML
SUBJECT_SAMPLE_FACTORS           	-	sample 3	Sample source:Cultured cells | Cell type:normal	Treatment=DMSO; RAW_FILE_NAME(Raw file name)=sample 3.mzML
SUBJECT_SAMPLE_FACTORS           	-	sample 4	Sample source:Cultured cells | Cell type:normal	Treatment=DMSO; RAW_FILE_NAME(Raw file name)=sample 4.mzML
SUBJECT_SAMPLE_FACTORS           	-	sample 5	Sample source:Cultured cells | Cell type:normal	Treatment=DMSO; RAW_FILE_NAME(Raw file name)=sample 5.mzML
SUBJECT_SAMPLE_FACTORS           	-	sample 6	Sample source:Cultured cells | Cell type:normal	Treatment=DMSO; RAW_FILE_NAME(Raw file name)=sample 6.mzML
SUBJECT_SAMPLE_FACTORS           	-	sample 7	Sample source:Cultured cells | Cell type:normal	Treatment=DMSO; RAW_FILE_NAME(Raw file name)=sample 7.mzML
SUBJECT_SAMPLE_FACTORS           	-	sample 8	Sample source:Cultured cells | Cell type:normal	Treatment=DMSO; RAW_FILE_NAME(Raw file name)=sample 8.mzML
SUBJECT_SAMPLE_FACTORS           	-	sample 9	Sample source:Cultured cells | Cell type:normal	Treatment=DMSO; RAW_FILE_NAME(Raw file name)=sample 9.mzML
SUBJECT_SAMPLE_FACTORS           	-	sample 10	Sample source:Cultured cells | Cell type:normal	Treatment=Erastin; RAW_FILE_NAME(Raw file name)=sample 10.mzML
SUBJECT_SAMPLE_FACTORS           	-	sample 11	Sample source:Cultured cells | Cell type:normal	Treatment=Erastin; RAW_FILE_NAME(Raw file name)=sample 11.mzML
SUBJECT_SAMPLE_FACTORS           	-	sample 12	Sample source:Cultured cells | Cell type:normal	Treatment=Erastin; RAW_FILE_NAME(Raw file name)=sample 12.mzML
SUBJECT_SAMPLE_FACTORS           	-	sample 13	Sample source:Cultured cells | Cell type:normal	Treatment=Erastin; RAW_FILE_NAME(Raw file name)=sample 13.mzML
SUBJECT_SAMPLE_FACTORS           	-	sample 14	Sample source:Cultured cells | Cell type:normal	Treatment=Erastin; RAW_FILE_NAME(Raw file name)=sample 14.mzML
SUBJECT_SAMPLE_FACTORS           	-	sample 15	Sample source:Cultured cells | Cell type:normal	Treatment=Erastin; RAW_FILE_NAME(Raw file name)=sample 15.mzML
SUBJECT_SAMPLE_FACTORS           	-	sample 16	Sample source:Cultured cells | Cell type:normal	Treatment=Erastin; RAW_FILE_NAME(Raw file name)=sample 16.mzML
SUBJECT_SAMPLE_FACTORS           	-	sample 17	Sample source:Cultured cells | Cell type:normal	Treatment=Erastin; RAW_FILE_NAME(Raw file name)=sample 17.mzML
SUBJECT_SAMPLE_FACTORS           	-	sample 18	Sample source:Cultured cells | Cell type:normal	Treatment=Erastin; RAW_FILE_NAME(Raw file name)=sample 18.mzML
SUBJECT_SAMPLE_FACTORS           	-	sample 19	Sample source:Cultured cells | Cell type:normal	Treatment=ConC; RAW_FILE_NAME(Raw file name)=sample 19.mzML
SUBJECT_SAMPLE_FACTORS           	-	sample 20	Sample source:Cultured cells | Cell type:normal	Treatment=ConC; RAW_FILE_NAME(Raw file name)=sample 20.mzML
SUBJECT_SAMPLE_FACTORS           	-	sample 21	Sample source:Cultured cells | Cell type:normal	Treatment=ConC; RAW_FILE_NAME(Raw file name)=sample 21.mzML
SUBJECT_SAMPLE_FACTORS           	-	sample 22	Sample source:Cultured cells | Cell type:normal	Treatment=ConC; RAW_FILE_NAME(Raw file name)=sample 22.mzML
SUBJECT_SAMPLE_FACTORS           	-	sample 23	Sample source:Cultured cells | Cell type:normal	Treatment=ConC; RAW_FILE_NAME(Raw file name)=sample 23.mzML
SUBJECT_SAMPLE_FACTORS           	-	sample 24	Sample source:Cultured cells | Cell type:normal	Treatment=ConC; RAW_FILE_NAME(Raw file name)=sample 24.mzML
SUBJECT_SAMPLE_FACTORS           	-	sample 25	Sample source:Cultured cells | Cell type:normal	Treatment=ConC; RAW_FILE_NAME(Raw file name)=sample 25.mzML
SUBJECT_SAMPLE_FACTORS           	-	sample 26	Sample source:Cultured cells | Cell type:normal	Treatment=ConC; RAW_FILE_NAME(Raw file name)=sample 26.mzML
SUBJECT_SAMPLE_FACTORS           	-	sample 27	Sample source:Cultured cells | Cell type:normal	Treatment=ConC; RAW_FILE_NAME(Raw file name)=sample 27.mzML
SUBJECT_SAMPLE_FACTORS           	-	sample 28	Sample source:Cultured cells | Cell type:normal	Treatment=Erastin+ConC; RAW_FILE_NAME(Raw file name)=sample 28.mzML
SUBJECT_SAMPLE_FACTORS           	-	sample 29	Sample source:Cultured cells | Cell type:normal	Treatment=Erastin+ConC; RAW_FILE_NAME(Raw file name)=sample 29.mzML
SUBJECT_SAMPLE_FACTORS           	-	sample 30	Sample source:Cultured cells | Cell type:normal	Treatment=Erastin+ConC; RAW_FILE_NAME(Raw file name)=sample 30.mzML
SUBJECT_SAMPLE_FACTORS           	-	sample 31	Sample source:Cultured cells | Cell type:normal	Treatment=Erastin+ConC; RAW_FILE_NAME(Raw file name)=sample 31.mzML
SUBJECT_SAMPLE_FACTORS           	-	sample 32	Sample source:Cultured cells | Cell type:normal	Treatment=Erastin+ConC; RAW_FILE_NAME(Raw file name)=sample 32.mzML
SUBJECT_SAMPLE_FACTORS           	-	sample 33	Sample source:Cultured cells | Cell type:normal	Treatment=Erastin+ConC; RAW_FILE_NAME(Raw file name)=sample 33.mzML
SUBJECT_SAMPLE_FACTORS           	-	sample 34	Sample source:Cultured cells | Cell type:normal	Treatment=Erastin+ConC; RAW_FILE_NAME(Raw file name)=sample 34.mzML
SUBJECT_SAMPLE_FACTORS           	-	sample 35	Sample source:Cultured cells | Cell type:normal	Treatment=Erastin+ConC; RAW_FILE_NAME(Raw file name)=sample 35.mzML
SUBJECT_SAMPLE_FACTORS           	-	sample 36	Sample source:Cultured cells | Cell type:normal	Treatment=Erastin+ConC; RAW_FILE_NAME(Raw file name)=sample 36.mzML
SUBJECT_SAMPLE_FACTORS           	-	sample 37	Sample source:Cultured cells | Cell type:senescent	Treatment=DMSO; RAW_FILE_NAME(Raw file name)=sample 37.mzML
SUBJECT_SAMPLE_FACTORS           	-	sample 38	Sample source:Cultured cells | Cell type:senescent	Treatment=DMSO; RAW_FILE_NAME(Raw file name)=sample 38.mzML
SUBJECT_SAMPLE_FACTORS           	-	sample 39	Sample source:Cultured cells | Cell type:senescent	Treatment=DMSO; RAW_FILE_NAME(Raw file name)=sample 39.mzML
SUBJECT_SAMPLE_FACTORS           	-	sample 40	Sample source:Cultured cells | Cell type:senescent	Treatment=DMSO; RAW_FILE_NAME(Raw file name)=sample 40.mzML
SUBJECT_SAMPLE_FACTORS           	-	sample 41	Sample source:Cultured cells | Cell type:senescent	Treatment=DMSO; RAW_FILE_NAME(Raw file name)=sample 41.mzML
SUBJECT_SAMPLE_FACTORS           	-	sample 42	Sample source:Cultured cells | Cell type:senescent	Treatment=DMSO; RAW_FILE_NAME(Raw file name)=sample 42.mzML
SUBJECT_SAMPLE_FACTORS           	-	sample 43	Sample source:Cultured cells | Cell type:senescent	Treatment=DMSO; RAW_FILE_NAME(Raw file name)=sample 43.mzML
SUBJECT_SAMPLE_FACTORS           	-	sample 44	Sample source:Cultured cells | Cell type:senescent	Treatment=DMSO; RAW_FILE_NAME(Raw file name)=sample 44.mzML
SUBJECT_SAMPLE_FACTORS           	-	sample 45	Sample source:Cultured cells | Cell type:senescent	Treatment=DMSO; RAW_FILE_NAME(Raw file name)=sample 45.mzML
SUBJECT_SAMPLE_FACTORS           	-	sample 46	Sample source:Cultured cells | Cell type:senescent	Treatment=Erastin; RAW_FILE_NAME(Raw file name)=sample 46.mzML
SUBJECT_SAMPLE_FACTORS           	-	sample 47	Sample source:Cultured cells | Cell type:senescent	Treatment=Erastin; RAW_FILE_NAME(Raw file name)=sample 47.mzML
SUBJECT_SAMPLE_FACTORS           	-	sample 48	Sample source:Cultured cells | Cell type:senescent	Treatment=Erastin; RAW_FILE_NAME(Raw file name)=sample 48.mzML
SUBJECT_SAMPLE_FACTORS           	-	sample 49	Sample source:Cultured cells | Cell type:senescent	Treatment=Erastin; RAW_FILE_NAME(Raw file name)=sample 49.mzML
SUBJECT_SAMPLE_FACTORS           	-	sample 50	Sample source:Cultured cells | Cell type:senescent	Treatment=Erastin; RAW_FILE_NAME(Raw file name)=sample 50.mzML
SUBJECT_SAMPLE_FACTORS           	-	sample 51	Sample source:Cultured cells | Cell type:senescent	Treatment=Erastin; RAW_FILE_NAME(Raw file name)=sample 51.mzML
SUBJECT_SAMPLE_FACTORS           	-	sample 52	Sample source:Cultured cells | Cell type:senescent	Treatment=Erastin; RAW_FILE_NAME(Raw file name)=sample 52.mzML
SUBJECT_SAMPLE_FACTORS           	-	sample 53	Sample source:Cultured cells | Cell type:senescent	Treatment=Erastin; RAW_FILE_NAME(Raw file name)=sample 53.mzML
SUBJECT_SAMPLE_FACTORS           	-	sample 54	Sample source:Cultured cells | Cell type:senescent	Treatment=Erastin; RAW_FILE_NAME(Raw file name)=sample 54.mzML
SUBJECT_SAMPLE_FACTORS           	-	sample 55	Sample source:Cultured cells | Cell type:senescent	Treatment=EN6; RAW_FILE_NAME(Raw file name)=sample 55.mzML
SUBJECT_SAMPLE_FACTORS           	-	sample 56	Sample source:Cultured cells | Cell type:senescent	Treatment=EN6; RAW_FILE_NAME(Raw file name)=sample 56.mzML
SUBJECT_SAMPLE_FACTORS           	-	sample 57	Sample source:Cultured cells | Cell type:senescent	Treatment=EN6; RAW_FILE_NAME(Raw file name)=sample 57.mzML
SUBJECT_SAMPLE_FACTORS           	-	sample 58	Sample source:Cultured cells | Cell type:senescent	Treatment=EN6; RAW_FILE_NAME(Raw file name)=sample 58.mzML
SUBJECT_SAMPLE_FACTORS           	-	sample 59	Sample source:Cultured cells | Cell type:senescent	Treatment=EN6; RAW_FILE_NAME(Raw file name)=sample 59.mzML
SUBJECT_SAMPLE_FACTORS           	-	sample 60	Sample source:Cultured cells | Cell type:senescent	Treatment=EN6; RAW_FILE_NAME(Raw file name)=sample 60.mzML
SUBJECT_SAMPLE_FACTORS           	-	sample 61	Sample source:Cultured cells | Cell type:senescent	Treatment=EN6; RAW_FILE_NAME(Raw file name)=sample 61.mzML
SUBJECT_SAMPLE_FACTORS           	-	sample 62	Sample source:Cultured cells | Cell type:senescent	Treatment=EN6; RAW_FILE_NAME(Raw file name)=sample 62.mzML
SUBJECT_SAMPLE_FACTORS           	-	sample 63	Sample source:Cultured cells | Cell type:senescent	Treatment=EN6; RAW_FILE_NAME(Raw file name)=sample 63.mzML
SUBJECT_SAMPLE_FACTORS           	-	sample 64	Sample source:Cultured cells | Cell type:senescent	Treatment=Erastin+EN6; RAW_FILE_NAME(Raw file name)=sample 64.mzML
SUBJECT_SAMPLE_FACTORS           	-	sample 65	Sample source:Cultured cells | Cell type:senescent	Treatment=Erastin+EN6; RAW_FILE_NAME(Raw file name)=sample 65.mzML
SUBJECT_SAMPLE_FACTORS           	-	sample 66	Sample source:Cultured cells | Cell type:senescent	Treatment=Erastin+EN6; RAW_FILE_NAME(Raw file name)=sample 66.mzML
SUBJECT_SAMPLE_FACTORS           	-	sample 67	Sample source:Cultured cells | Cell type:senescent	Treatment=Erastin+EN6; RAW_FILE_NAME(Raw file name)=sample 67.mzML
SUBJECT_SAMPLE_FACTORS           	-	sample 68	Sample source:Cultured cells | Cell type:senescent	Treatment=Erastin+EN6; RAW_FILE_NAME(Raw file name)=sample 68.mzML
SUBJECT_SAMPLE_FACTORS           	-	sample 69	Sample source:Cultured cells | Cell type:senescent	Treatment=Erastin+EN6; RAW_FILE_NAME(Raw file name)=sample 69.mzML
SUBJECT_SAMPLE_FACTORS           	-	sample 70	Sample source:Cultured cells | Cell type:senescent	Treatment=Erastin+EN6; RAW_FILE_NAME(Raw file name)=sample 70.mzML
SUBJECT_SAMPLE_FACTORS           	-	sample 71	Sample source:Cultured cells | Cell type:senescent	Treatment=Erastin+EN6; RAW_FILE_NAME(Raw file name)=sample 71.mzML
SUBJECT_SAMPLE_FACTORS           	-	sample 72	Sample source:Cultured cells | Cell type:senescent	Treatment=Erastin+EN6; RAW_FILE_NAME(Raw file name)=sample 72.mzML
#COLLECTION
CO:COLLECTION_SUMMARY            	Human normal fibroblasts (TIG-3, JCRB0506) was obtained from the JCRB cell bank.
CO:COLLECTION_SUMMARY            	Cells used in this experiment were grown in Dulbecco's Modified Eagle Medium
CO:COLLECTION_SUMMARY            	(DMEM; Nacalai Tesque, 16971-55) containing 10% fetal bovine serum (FBS) (Gibco,
CO:COLLECTION_SUMMARY            	10437-028) and penicillin/streptomycin (Fujifilm Wako Chemicals, 168-23191). For
CO:COLLECTION_SUMMARY            	induction of cellular senescence by X-irradiation, TIG-3 cells were seeded at a
CO:COLLECTION_SUMMARY            	density of 2500 cells/cm2 and cultured for 11 days after irradiating TIG-3 cells
CO:COLLECTION_SUMMARY            	with 15 Gy of X-rays using CP-160 (Faxitron X-ray Corporation).
CO:SAMPLE_TYPE                   	Fibroblasts
#TREATMENT
TR:TREATMENT_SUMMARY             	To induce ferroptosis in cells, cells were cultured in medium containing erastin
TR:TREATMENT_SUMMARY             	(Selleck, S7242) for 24 h. Ferroptosis inducer; erastin was administered to
TR:TREATMENT_SUMMARY             	cells with erastin and cultured for 24 hours. Concanamycin C (Adipogen Life
TR:TREATMENT_SUMMARY             	Sciences, BVT-0254-C100) or EN6 (Selleck, S6650), as compounds that alter
TR:TREATMENT_SUMMARY             	lysosomal pH, were also coadministered with erastin and cells were incubated for
TR:TREATMENT_SUMMARY             	24 hours.
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	Lipids and oxidized lipids were extracted by adding 1.0 mL of a 1:1 (v/v)
SP:SAMPLEPREP_SUMMARY            	solution of 1-butanol and methanol, containing 100 µM butylated hydroxytoluene
SP:SAMPLEPREP_SUMMARY            	(BHT) and 100 µM ethylenediaminetetraacetic acid (EDTA), to the cell pellets.
SP:SAMPLEPREP_SUMMARY            	The mixture was vortexed for 30 s, sonicated for 5 min in an ice-cooled sonic
SP:SAMPLEPREP_SUMMARY            	bath, and then centrifuged at 16,000 × g for 10 minutes at 4 °C. The resultant
SP:SAMPLEPREP_SUMMARY            	supernatant was transferred into a 0.2 mL glass insert equipped with a
SP:SAMPLEPREP_SUMMARY            	Teflon-lined cap for subsequent LC ESI-MS analysis.
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_TYPE           	Reversed phase
CH:INSTRUMENT_NAME               	Thermo Dionex Ultimate 3000
CH:COLUMN_NAME                   	Thermo Accucore C18 (150 x 2.1mm,2.6um)
CH:SOLVENT_A                     	50% acetonitrile/50% water; 0.1% formic acid; 10 mM ammonium formate
CH:SOLVENT_B                     	10% acetonitrile/88% isopropyl alcohol/2% water; 0.02% formic acid; 2 mM
CH:SOLVENT_B                     	ammonium formate
CH:FLOW_GRADIENT                 	From 65:35 (A:B) at onset to 40:60 over 0 to 4 min, shifting to 15:85 from 4 to
CH:FLOW_GRADIENT                 	12 minutes, then to 0:100 from 12 to 21 minutes, maintained at 0:100 from 21 to
CH:FLOW_GRADIENT                 	24 min, reverted to 65:35 over 24 to 24.1 min, and finally to 100:0 from 24.1 to
CH:FLOW_GRADIENT                 	28 min
CH:FLOW_RATE                     	0.4 mL/min
CH:COLUMN_TEMPERATURE            	35°C
#ANALYSIS
AN:ANALYSIS_TYPE                 	MS
#MS
MS:INSTRUMENT_NAME               	Thermo Orbitrap Exploris 240
MS:INSTRUMENT_TYPE               	Orbitrap
MS:MS_TYPE                       	ESI
MS:ION_MODE                      	POSITIVE
MS:MS_COMMENTS                   	The Orbitrap Exploris mass spectrometer was operated in both ESI positive and
MS:MS_COMMENTS                   	negative ion modes. A full mass scan (m/z 250–1200) was followed by three
MS:MS_COMMENTS                   	rapid data-dependent MS/MS scans with resolutions of 70,000 and 17,500. The
MS:MS_COMMENTS                   	instrument settings included an automatic gain control target of 1 × 106 ions
MS:MS_COMMENTS                   	and a maximum ion injection time of 100 ms. Ion source adjustments were set with
MS:MS_COMMENTS                   	a spray voltage of 3 kV, a transfer tube temperature of 285°C, an S-Lens level
MS:MS_COMMENTS                   	of 45, a heater temperature of 370°C, a sheath gas flow rate of 60, and an
MS:MS_COMMENTS                   	auxiliary gas flow rate of 20.
#MS_METABOLITE_DATA
MS_METABOLITE_DATA:UNITS	oxidized PL/non-oxidized PL, %
MS_METABOLITE_DATA_START
Samples	sample 1	sample 2	sample 3	sample 4	sample 5	sample 6	sample 7	sample 8	sample 9	sample 10	sample 11	sample 12	sample 13	sample 14	sample 15	sample 16	sample 17	sample 18	sample 19	sample 20	sample 21	sample 22	sample 23	sample 24	sample 25	sample 26	sample 27	sample 28	sample 29	sample 30	sample 31	sample 32	sample 33	sample 34	sample 35	sample 36	sample 37	sample 38	sample 39	sample 40	sample 41	sample 42	sample 43	sample 44	sample 45	sample 46	sample 47	sample 48	sample 49	sample 50	sample 51	sample 52	sample 53	sample 54	sample 55	sample 56	sample 57	sample 58	sample 59	sample 60	sample 61	sample 62	sample 63	sample 64	sample 65	sample 66	sample 67	sample 68	sample 69	sample 70	sample 71	sample 72
Factors	Sample source:Cultured cells | Cell type:normal	Sample source:Cultured cells | Cell type:normal	Sample source:Cultured cells | Cell type:normal	Sample source:Cultured cells | Cell type:normal	Sample source:Cultured cells | Cell type:normal	Sample source:Cultured cells | Cell type:normal	Sample source:Cultured cells | Cell type:normal	Sample source:Cultured cells | Cell type:normal	Sample source:Cultured cells | Cell type:normal	Sample source:Cultured cells | Cell type:normal	Sample source:Cultured cells | Cell type:normal	Sample source:Cultured cells | Cell type:normal	Sample source:Cultured cells | Cell type:normal	Sample source:Cultured cells | Cell type:normal	Sample source:Cultured cells | Cell type:normal	Sample source:Cultured cells | Cell type:normal	Sample source:Cultured cells | Cell type:normal	Sample source:Cultured cells | Cell type:normal	Sample source:Cultured cells | Cell type:normal	Sample source:Cultured cells | Cell type:normal	Sample source:Cultured cells | Cell type:normal	Sample source:Cultured cells | Cell type:normal	Sample source:Cultured cells | Cell type:normal	Sample source:Cultured cells | Cell type:normal	Sample source:Cultured cells | Cell type:normal	Sample source:Cultured cells | Cell type:normal	Sample source:Cultured cells | Cell type:normal	Sample source:Cultured cells | Cell type:normal	Sample source:Cultured cells | Cell type:normal	Sample source:Cultured cells | Cell type:normal	Sample source:Cultured cells | Cell type:normal	Sample source:Cultured cells | Cell type:normal	Sample source:Cultured cells | Cell type:normal	Sample source:Cultured cells | Cell type:normal	Sample source:Cultured cells | Cell type:normal	Sample source:Cultured cells | Cell type:normal	Sample source:Cultured cells | Cell type:senescent	Sample source:Cultured cells | Cell type:senescent	Sample source:Cultured cells | Cell type:senescent	Sample source:Cultured cells | Cell type:senescent	Sample source:Cultured cells | Cell type:senescent	Sample source:Cultured cells | Cell type:senescent	Sample source:Cultured cells | Cell type:senescent	Sample source:Cultured cells | Cell type:senescent	Sample source:Cultured cells | Cell type:senescent	Sample source:Cultured cells | Cell type:senescent	Sample source:Cultured cells | Cell type:senescent	Sample source:Cultured cells | Cell type:senescent	Sample source:Cultured cells | Cell type:senescent	Sample source:Cultured cells | Cell type:senescent	Sample source:Cultured cells | Cell type:senescent	Sample source:Cultured cells | Cell type:senescent	Sample source:Cultured cells | Cell type:senescent	Sample source:Cultured cells | Cell type:senescent	Sample source:Cultured cells | Cell type:senescent	Sample source:Cultured cells | Cell type:senescent	Sample source:Cultured cells | Cell type:senescent	Sample source:Cultured cells | Cell type:senescent	Sample source:Cultured cells | Cell type:senescent	Sample source:Cultured cells | Cell type:senescent	Sample source:Cultured cells | Cell type:senescent	Sample source:Cultured cells | Cell type:senescent	Sample source:Cultured cells | Cell type:senescent	Sample source:Cultured cells | Cell type:senescent	Sample source:Cultured cells | Cell type:senescent	Sample source:Cultured cells | Cell type:senescent	Sample source:Cultured cells | Cell type:senescent	Sample source:Cultured cells | Cell type:senescent	Sample source:Cultured cells | Cell type:senescent	Sample source:Cultured cells | Cell type:senescent	Sample source:Cultured cells | Cell type:senescent	Sample source:Cultured cells | Cell type:senescent
PC38:4;O	0.50194	0.41893	0.44478	0.36334	0.46631	0.45098	0.47140	0.42494	0.33088	0.44978	0.29710	0.45199	0.43725	0.34860	0.45714	0.37746	0.67483	0.41332	0.98746	0.49573	0.87010	0.69349	0.71425	1.02481	0.87045	0.94202	1.18629	0.34651	0.31218	0.78303	0.36034	0.43620	0.41732	0.40698	0.48899	0.31554	0.501939	0.418930169	0.44478	0.36334	0.46631	0.45098	0.47140	0.42494	0.33088	0.44978	0.29710	0.45199	0.43725	0.34860	0.45714	0.37746	0.67483	0.41332	0.98746	0.49573	0.87010	0.69349	0.71425	1.02481	0.87045	0.94202	1.18629	0.34651	0.31218	0.78303	0.36034	0.43620	0.41732	0.40698	0.48899	0.31554
PC38:4;O2	0.52573	0.44781	0.51109	0.31786	0.50945	0.49404	0.49398	0.45283	0.33510	0.52256	0.30043	0.52316	0.47694	0.35720	0.51502	0.38155	0.83031	0.43444	1.22101	0.53543	1.20461	0.72509	0.72293	1.24240	0.84855	1.11252	1.45073	0.34570	0.31214	0.94415	0.37122	0.48552	0.48263	0.43524	0.54599	0.31388	0.525725779	0.447805525	0.51109	0.31786	0.50945	0.49404	0.49398	0.45283	0.33510	0.52256	0.30043	0.52316	0.47694	0.35720	0.51502	0.38155	0.83031	0.43444	1.22101	0.53543	1.20461	0.72509	0.72293	1.24240	0.84855	1.11252	1.45073	0.34570	0.31214	0.94415	0.37122	0.48552	0.48263	0.43524	0.54599	0.31388
PC38:4;O3	0.07902	0.05309	0.05542	0.03817	0.01761	0.05414	0.06787	0.05479	0.03530	0.06094	0.03373	0.05327	0.05799	0.03183	0.05375	0.03619	0.09320	0.04220	0.44253	0.26030	0.39963	0.26947	0.27528	0.24342	0.14634	0.24067	0.27873	0.03347	0.02791	0.10390	0.03827	0.05359	0.05195	0.05567	0.06408	0.02850	0.079024436	0.053094962	0.05542	0.03817	0.01761	0.05414	0.06787	0.05479	0.03530	0.06094	0.03373	0.05327	0.05799	0.03183	0.05375	0.03619	0.09320	0.04220	0.44253	0.26030	0.39963	0.26947	0.27528	0.24342	0.14634	0.24067	0.27873	0.03347	0.02791	0.10390	0.03827	0.05359	0.05195	0.05567	0.06408	0.02850
PC36:4;O	0.45321	0.39537	0.44640	0.36091	0.44326	0.43924	0.46410	0.41548	0.34161	0.48090	0.34255	0.45807	0.44023	0.35476	0.46227	0.36739	0.60581	0.42498	1.03482	0.51374	0.92802	0.65633	0.67207	0.82247	0.66199	0.82280	0.92912	0.32333	0.30139	0.58927	0.33610	0.40320	0.38532	0.38990	0.41865	0.29546	0.453208685	0.395372164	0.446398781	0.360914325	0.443258816	0.439238015	0.464104718	0.415483902	0.341606738	0.480899824	0.342547264	0.458070017	0.440229395	0.354763294	0.46227426	0.367392546	0.605814061	0.42498478	1.034818399	0.513736216	0.928017604	0.656332573	0.672066957	0.822473682	0.661991092	0.822799371	0.929117758	0.323334136	0.30139403	0.589273977	0.336099423	0.403201169	0.385322009	0.389899104	0.418650802	0.295464111
PC36:4;O2	0.15127	0.13711	0.15335	0.08968	0.15482	0.15088	0.15160	0.14386	0.10403	0.16944	0.10132	0.16575	0.14781	0.11878	0.01619	0.10862	0.21665	0.13649	0.53528	0.27688	0.51242	0.28786	0.27899	0.39403	0.26822	0.36342	0.46409	0.10056	0.08879	0.02187	0.10696	0.14894	0.14481	0.13404	0.15814	0.09489	0.151273881	0.137106327	0.153353429	0.0896784	0.154821622	0.150881496	0.151598532	0.143863158	0.104030665	0.169438722	0.101322554	0.165746791	0.147812637	0.118778686	0.016186288	0.108619963	0.216645773	0.136489519	0.535284238	0.276878029	0.512415405	0.28785704	0.278987436	0.394028324	0.2682228	0.363416347	0.464085722	0.100555923	0.088790633	0.021872538	0.106957829	0.148942355	0.144806208	0.134042685	0.158136509	0.094894115
PC36:4;O3	0.00418	0.00142	0.00028	0.00133	0.00131	0.00074	0.00180	0.00111	0.00055	0.00232	0.00000	0.00124	0.00000	0.00000	0.00000	0.00225	0.00244	0.00079	0.18313	0.14018	0.14848	0.10684	0.09919	0.05146	0.03421	0.05604	0.06613	0.00084	0.00043	0.00087	0.00199	0.00085	0.00061	0.00120	0.00088	0.00088	0.004184401	0.001423504	0.000284113	0.001328556	0.001306688	0.000735224	0.001799928	0.001111705	0.000553153	0.002319603	0	0.001239398	0	0	0	0.002245402	0.002443374	0.000788306	0.183134118	0.140176688	0.148477538	0.106835961	0.099194036	0.051463456	0.034208129	0.056039717	0.066130502	0.0008408	0.000426248	0.000870215	0.001991891	0.000847822	0.000611067	0.00120036	0.00088369	0.000875441
MS_METABOLITE_DATA_END
#METABOLITES
METABOLITES_START
metabolite_name
PC38:4;O
PC38:4;O2
PC38:4;O3
PC36:4;O
PC36:4;O2
PC36:4;O3
METABOLITES_END
#END