#METABOLOMICS WORKBENCH engelmab_20230926_074954 DATATRACK_ID:4335 STUDY_ID:ST002914 ANALYSIS_ID:AN004784 PROJECT_ID:PR001812
VERSION             	1
CREATED_ON             	October 4, 2023, 10:58 am
#PROJECT
PR:PROJECT_TITLE                 	Untangling the Dynamics of Lysine Acetylation and Phosphorylation in
PR:PROJECT_TITLE                 	Adipogenesis in the Established Human and Mouse Adipocyte Cell Lines SGBS and
PR:PROJECT_TITLE                 	3T3L1
PR:PROJECT_SUMMARY               	Obesity is one of the most pressing global public health challenges of our time
PR:PROJECT_SUMMARY               	with yet increasing prevalence. Characterized by enlarged and dysfunctional
PR:PROJECT_SUMMARY               	adipose tissue, it is often associated with the development of metabolic or
PR:PROJECT_SUMMARY               	cardiovascular comorbidities. A sound understanding of the processes underlying
PR:PROJECT_SUMMARY               	adipogenesis, the differentiation of a preadipocyte into a mature and
PR:PROJECT_SUMMARY               	lipid-laden adipocyte, provide the basic knowledge for future research on the
PR:PROJECT_SUMMARY               	causes and consequences of obesity. The tricarboxylic acid cycle represents the
PR:PROJECT_SUMMARY               	central metabolic hub, that provides energy equivalents, metabolic building
PR:PROJECT_SUMMARY               	blocks and critical intermediates such as acetyl-CoA as precursor for protein
PR:PROJECT_SUMMARY               	acetylation and de novo lipogenesis. Stable cell lines are an integral part of
PR:PROJECT_SUMMARY               	research into the development and physiology of adipocytes in health and disease
PR:PROJECT_SUMMARY               	and various models have been used to date. In this study we demonstrated the
PR:PROJECT_SUMMARY               	vivid temporal dynamics of central carbon metabolites during the differentiation
PR:PROJECT_SUMMARY               	of SGBS and 3T3-L1 adipocytes. Detected metabolite levels showed distinct
PR:PROJECT_SUMMARY               	temporal profiles, partly with cell-line specificities.
PR:INSTITUTE                     	Helmholtz Centre for Environmental Research
PR:DEPARTMENT                    	Molecular Systems Biology
PR:LAST_NAME                     	Engelmann
PR:FIRST_NAME                    	Beatrice
PR:ADDRESS                       	Permoserstraße 15, Leipzipg, Saxony, 03418, Germany
PR:EMAIL                         	beatrice.engelmann@ufz.de
PR:PHONE                         	00493412351099
#STUDY
ST:STUDY_TITLE                   	Untangling the Dynamics of Lysine Acetylation and Phosphorylation in
ST:STUDY_TITLE                   	Adipogenesis in the Established Human and Mouse Adipocyte Cell Lines SGBS and
ST:STUDY_TITLE                   	3T3L1
ST:STUDY_SUMMARY                 	Obesity is one of the most pressing global public health challenges of our time
ST:STUDY_SUMMARY                 	with yet increasing prevalence. Characterized by enlarged and dysfunctional
ST:STUDY_SUMMARY                 	adipose tissue, it is often associated with the development of metabolic or
ST:STUDY_SUMMARY                 	cardiovascular comorbidities. A sound understanding of the processes underlying
ST:STUDY_SUMMARY                 	adipogenesis, the differentiation of a preadipocyte into a mature and lipid
ST:STUDY_SUMMARY                 	laden adipocyte, provide the basic knowledge for future research on the causes
ST:STUDY_SUMMARY                 	and consequences of obesity. The tricarboxylic acid cycle represents the central
ST:STUDY_SUMMARY                 	metabolic hub, that provides energy equivalents, metabolic building blocks and
ST:STUDY_SUMMARY                 	critical intermediates such as acetyl-CoA as precursor for protein acetylation
ST:STUDY_SUMMARY                 	and de novo lipogenesis. Stable cell lines are an integral part of research into
ST:STUDY_SUMMARY                 	the development and physiology of adipocytes in health and disease and various
ST:STUDY_SUMMARY                 	models have been used to date. In this study we demonstrated the vivid temporal
ST:STUDY_SUMMARY                 	dynamics of central carbon metabolites during the differentiation of SGBS and
ST:STUDY_SUMMARY                 	3T3-L1 adipocytes. Detected metabolite levels showed distinct temporal profiles,
ST:STUDY_SUMMARY                 	partly with cell-line specificities.
ST:INSTITUTE                     	Helmholtz Centre for Environmental Research
ST:DEPARTMENT                    	Molecular Systems Biology
ST:LAST_NAME                     	Engelmann
ST:FIRST_NAME                    	Beatrice
ST:ADDRESS                       	Permoserstraße 15, Leipzipg, Saxony, 03418, Germany
ST:EMAIL                         	beatrice.engelmann@ufz.de
ST:PHONE                         	00493412351099
#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           	-	SGBS_ctrl_d0_1	Treatment:d0 | Replicate:1 | Species:Homo sapiens	RAW_FILE_NAME=221207_AA_acetylCoA_differentiation_SGBS_144_Alix.wiff
SUBJECT_SAMPLE_FACTORS           	-	SGBS_ctrl_d0_2	Treatment:d0 | Replicate:2 | Species:Homo sapiens	RAW_FILE_NAME=221206_AA_acetylCoA_differentiation_SGBS_144_Alix.wiff
SUBJECT_SAMPLE_FACTORS           	-	SGBS_ctrl_d0_3	Treatment:d0 | Replicate:3 | Species:Homo sapiens	RAW_FILE_NAME=221206_AA_acetylCoA_differentiation_SGBS_144_Alix.wiff
SUBJECT_SAMPLE_FACTORS           	-	SGBS_ctrl_d0_4	Treatment:d0 | Replicate:4 | Species:Homo sapiens	RAW_FILE_NAME=221206_AA_acetylCoA_differentiation_SGBS_144_Alix.wiff
SUBJECT_SAMPLE_FACTORS           	-	SGBS_ctrl_d0_5	Treatment:d0 | Replicate:5 | Species:Homo sapiens	RAW_FILE_NAME=221206_AA_acetylCoA_differentiation_SGBS_144_Alix.wiff
SUBJECT_SAMPLE_FACTORS           	-	SGBS_ctrl_d0_6	Treatment:d0 | Replicate:6 | Species:Homo sapiens	RAW_FILE_NAME=221206_AA_acetylCoA_differentiation_SGBS_144_Alix.wiff
SUBJECT_SAMPLE_FACTORS           	-	SGBS_ctrl_d3_1	Treatment:d3 | Replicate:1 | Species:Homo sapiens	RAW_FILE_NAME=221207_AA_acetylCoA_differentiation_SGBS_144_Alix.wiff
SUBJECT_SAMPLE_FACTORS           	-	SGBS_ctrl_d3_2	Treatment:d3 | Replicate:2 | Species:Homo sapiens	RAW_FILE_NAME=221207_AA_acetylCoA_differentiation_SGBS_144_Alix.wiff
SUBJECT_SAMPLE_FACTORS           	-	SGBS_ctrl_d3_3	Treatment:d3 | Replicate:3 | Species:Homo sapiens	RAW_FILE_NAME=221206_AA_acetylCoA_differentiation_SGBS_144_Alix.wiff
SUBJECT_SAMPLE_FACTORS           	-	SGBS_ctrl_d3_4	Treatment:d3 | Replicate:4 | Species:Homo sapiens	RAW_FILE_NAME=221207_AA_acetylCoA_differentiation_SGBS_144_Alix.wiff
SUBJECT_SAMPLE_FACTORS           	-	SGBS_ctrl_d3_5	Treatment:d3 | Replicate:5 | Species:Homo sapiens	RAW_FILE_NAME=221206_AA_acetylCoA_differentiation_SGBS_144_Alix.wiff
SUBJECT_SAMPLE_FACTORS           	-	SGBS_ctrl_d3_6	Treatment:d3 | Replicate:6 | Species:Homo sapiens	RAW_FILE_NAME=221207_AA_acetylCoA_differentiation_SGBS_144_Alix.wiff
SUBJECT_SAMPLE_FACTORS           	-	SGBS_ctrl_d6_1	Treatment:d6 | Replicate:1 | Species:Homo sapiens	RAW_FILE_NAME=221207_AA_acetylCoA_differentiation_SGBS_144_Alix.wiff
SUBJECT_SAMPLE_FACTORS           	-	SGBS_ctrl_d6_2	Treatment:d6 | Replicate:2 | Species:Homo sapiens	RAW_FILE_NAME=221207_AA_acetylCoA_differentiation_SGBS_144_Alix.wiff
SUBJECT_SAMPLE_FACTORS           	-	SGBS_ctrl_d6_3	Treatment:d6 | Replicate:3 | Species:Homo sapiens	RAW_FILE_NAME=221207_AA_acetylCoA_differentiation_SGBS_144_Alix.wiff
SUBJECT_SAMPLE_FACTORS           	-	SGBS_ctrl_d6_4	Treatment:d6 | Replicate:4 | Species:Homo sapiens	RAW_FILE_NAME=221207_AA_acetylCoA_differentiation_SGBS_144_Alix.wiff
SUBJECT_SAMPLE_FACTORS           	-	SGBS_ctrl_d6_5	Treatment:d6 | Replicate:5 | Species:Homo sapiens	RAW_FILE_NAME=221206_AA_acetylCoA_differentiation_SGBS_144_Alix.wiff
SUBJECT_SAMPLE_FACTORS           	-	SGBS_ctrl_d6_6	Treatment:d6 | Replicate:6 | Species:Homo sapiens	RAW_FILE_NAME=221206_AA_acetylCoA_differentiation_SGBS_144_Alix.wiff
SUBJECT_SAMPLE_FACTORS           	-	SGBS_ctrl_d9_1	Treatment:d9 | Replicate:1 | Species:Homo sapiens	RAW_FILE_NAME=221207_AA_acetylCoA_differentiation_SGBS_144_Alix.wiff
SUBJECT_SAMPLE_FACTORS           	-	SGBS_ctrl_d9_2	Treatment:d9 | Replicate:2 | Species:Homo sapiens	RAW_FILE_NAME=221206_AA_acetylCoA_differentiation_SGBS_144_Alix.wiff
SUBJECT_SAMPLE_FACTORS           	-	SGBS_ctrl_d9_3	Treatment:d9 | Replicate:3 | Species:Homo sapiens	RAW_FILE_NAME=221207_AA_acetylCoA_differentiation_SGBS_144_Alix.wiff
SUBJECT_SAMPLE_FACTORS           	-	SGBS_ctrl_d9_4	Treatment:d9 | Replicate:4 | Species:Homo sapiens	RAW_FILE_NAME=221206_AA_acetylCoA_differentiation_SGBS_144_Alix.wiff
SUBJECT_SAMPLE_FACTORS           	-	SGBS_ctrl_d9_5	Treatment:d9 | Replicate:5 | Species:Homo sapiens	RAW_FILE_NAME=221206_AA_acetylCoA_differentiation_SGBS_144_Alix.wiff
SUBJECT_SAMPLE_FACTORS           	-	SGBS_ctrl_d9_6	Treatment:d9 | Replicate:6 | Species:Homo sapiens	RAW_FILE_NAME=221207_AA_acetylCoA_differentiation_SGBS_144_Alix.wiff
SUBJECT_SAMPLE_FACTORS           	-	SGBS_ctrl_d12_1	Treatment:d12 | Replicate:1 | Species:Homo sapiens	RAW_FILE_NAME=221206_AA_acetylCoA_differentiation_SGBS_144_Alix.wiff
SUBJECT_SAMPLE_FACTORS           	-	SGBS_ctrl_d12_2	Treatment:d12 | Replicate:2 | Species:Homo sapiens	RAW_FILE_NAME=221206_AA_acetylCoA_differentiation_SGBS_144_Alix.wiff
SUBJECT_SAMPLE_FACTORS           	-	SGBS_ctrl_d12_3	Treatment:d12 | Replicate:3 | Species:Homo sapiens	RAW_FILE_NAME=221206_AA_acetylCoA_differentiation_SGBS_144_Alix.wiff
SUBJECT_SAMPLE_FACTORS           	-	SGBS_ctrl_d12_4	Treatment:d12 | Replicate:4 | Species:Homo sapiens	RAW_FILE_NAME=221207_AA_acetylCoA_differentiation_SGBS_144_Alix.wiff
SUBJECT_SAMPLE_FACTORS           	-	SGBS_ctrl_d12_5	Treatment:d12 | Replicate:5 | Species:Homo sapiens	RAW_FILE_NAME=221207_AA_acetylCoA_differentiation_SGBS_144_Alix.wiff
SUBJECT_SAMPLE_FACTORS           	-	SGBS_ctrl_d12_6	Treatment:d12 | Replicate:6 | Species:Homo sapiens	RAW_FILE_NAME=221206_AA_acetylCoA_differentiation_SGBS_144_Alix.wiff
SUBJECT_SAMPLE_FACTORS           	-	3T3L1_ctrl_d0_1	Treatment:d0 | Replicate:1 | Species:Mus musculus	RAW_FILE_NAME=221125_AceCoA_diff_3T3L1_Alix_Set2.wiff
SUBJECT_SAMPLE_FACTORS           	-	3T3L1_ctrl_d0_2	Treatment:d0 | Replicate:2 | Species:Mus musculus	RAW_FILE_NAME=221125_AceCoA_diff_3T3L1_Alix_Set2.wiff
SUBJECT_SAMPLE_FACTORS           	-	3T3L1_ctrl_d0_3	Treatment:d0 | Replicate:3 | Species:Mus musculus	RAW_FILE_NAME=221123_AceCoA_diff_3T3L1_Alix.wiff
SUBJECT_SAMPLE_FACTORS           	-	3T3L1_ctrl_d0_4	Treatment:d0 | Replicate:4 | Species:Mus musculus	RAW_FILE_NAME=221125_AceCoA_diff_3T3L1_Alix_Set2.wiff
SUBJECT_SAMPLE_FACTORS           	-	3T3L1_ctrl_d0_5	Treatment:d0 | Replicate:5 | Species:Mus musculus	RAW_FILE_NAME=221125_AceCoA_diff_3T3L1_Alix_Set2.wiff
SUBJECT_SAMPLE_FACTORS           	-	3T3L1_ctrl_d0_6	Treatment:d0 | Replicate:6 | Species:Mus musculus	RAW_FILE_NAME=221123_AceCoA_diff_3T3L1_Alix.wiff
SUBJECT_SAMPLE_FACTORS           	-	3T3L1_ctrl_d3_1	Treatment:d3 | Replicate:1 | Species:Mus musculus	RAW_FILE_NAME=221125_AceCoA_diff_3T3L1_Alix_Set2.wiff
SUBJECT_SAMPLE_FACTORS           	-	3T3L1_ctrl_d3_2	Treatment:d3 | Replicate:2 | Species:Mus musculus	RAW_FILE_NAME=221125_AceCoA_diff_3T3L1_Alix_Set2.wiff
SUBJECT_SAMPLE_FACTORS           	-	3T3L1_ctrl_d3_3	Treatment:d3 | Replicate:3 | Species:Mus musculus	RAW_FILE_NAME=221125_AceCoA_diff_3T3L1_Alix_Set2.wiff
SUBJECT_SAMPLE_FACTORS           	-	3T3L1_ctrl_d3_4	Treatment:d3 | Replicate:4 | Species:Mus musculus	RAW_FILE_NAME=221125_AceCoA_diff_3T3L1_Alix_Set2.wiff
SUBJECT_SAMPLE_FACTORS           	-	3T3L1_ctrl_d3_5	Treatment:d3 | Replicate:5 | Species:Mus musculus	RAW_FILE_NAME=221125_AceCoA_diff_3T3L1_Alix_Set2.wiff
SUBJECT_SAMPLE_FACTORS           	-	3T3L1_ctrl_d3_6	Treatment:d3 | Replicate:6 | Species:Mus musculus	RAW_FILE_NAME=221125_AceCoA_diff_3T3L1_Alix_Set2.wiff
SUBJECT_SAMPLE_FACTORS           	-	3T3L1_ctrl_d5_1	Treatment:d5 | Replicate:1 | Species:Mus musculus	RAW_FILE_NAME=221123_AceCoA_diff_3T3L1_Alix.wiff
SUBJECT_SAMPLE_FACTORS           	-	3T3L1_ctrl_d5_2	Treatment:d5 | Replicate:2 | Species:Mus musculus	RAW_FILE_NAME=221125_AceCoA_diff_3T3L1_Alix_Set2.wiff
SUBJECT_SAMPLE_FACTORS           	-	3T3L1_ctrl_d5_3	Treatment:d5 | Replicate:3 | Species:Mus musculus	RAW_FILE_NAME=221123_AceCoA_diff_3T3L1_Alix.wiff
SUBJECT_SAMPLE_FACTORS           	-	3T3L1_ctrl_d5_4	Treatment:d5 | Replicate:4 | Species:Mus musculus	RAW_FILE_NAME=221123_AceCoA_diff_3T3L1_Alix.wiff
SUBJECT_SAMPLE_FACTORS           	-	3T3L1_ctrl_d5_5	Treatment:d5 | Replicate:5 | Species:Mus musculus	RAW_FILE_NAME=221125_AceCoA_diff_3T3L1_Alix_Set2.wiff
SUBJECT_SAMPLE_FACTORS           	-	3T3L1_ctrl_d5_6	Treatment:d5 | Replicate:6 | Species:Mus musculus	RAW_FILE_NAME=221125_AceCoA_diff_3T3L1_Alix_Set2.wiff
SUBJECT_SAMPLE_FACTORS           	-	3T3L1_ctrl_d7_1	Treatment:d7 | Replicate:1 | Species:Mus musculus	RAW_FILE_NAME=221125_AceCoA_diff_3T3L1_Alix_Set2.wiff
SUBJECT_SAMPLE_FACTORS           	-	3T3L1_ctrl_d7_2	Treatment:d7 | Replicate:2 | Species:Mus musculus	RAW_FILE_NAME=221125_AceCoA_diff_3T3L1_Alix_Set2.wiff
SUBJECT_SAMPLE_FACTORS           	-	3T3L1_ctrl_d7_3	Treatment:d7 | Replicate:3 | Species:Mus musculus	RAW_FILE_NAME=221125_AceCoA_diff_3T3L1_Alix_Set2.wiff
SUBJECT_SAMPLE_FACTORS           	-	3T3L1_ctrl_d7_4	Treatment:d7 | Replicate:4 | Species:Mus musculus	RAW_FILE_NAME=221123_AceCoA_diff_3T3L1_Alix.wiff
SUBJECT_SAMPLE_FACTORS           	-	3T3L1_ctrl_d7_5	Treatment:d7 | Replicate:5 | Species:Mus musculus	RAW_FILE_NAME=221125_AceCoA_diff_3T3L1_Alix_Set2.wiff
SUBJECT_SAMPLE_FACTORS           	-	3T3L1_ctrl_d7_6	Treatment:d7 | Replicate:6 | Species:Mus musculus	RAW_FILE_NAME=221123_AceCoA_diff_3T3L1_Alix.wiff
SUBJECT_SAMPLE_FACTORS           	-	3T3L1_ctrl_d10_1	Treatment:d1 | Replicate:1 | Species:Mus musculus	RAW_FILE_NAME=221123_AceCoA_diff_3T3L1_Alix.wiff
SUBJECT_SAMPLE_FACTORS           	-	3T3L1_ctrl_d10_2	Treatment:d1 | Replicate:2 | Species:Mus musculus	RAW_FILE_NAME=221125_AceCoA_diff_3T3L1_Alix_Set2.wiff
SUBJECT_SAMPLE_FACTORS           	-	3T3L1_ctrl_d10_3	Treatment:d1 | Replicate:3 | Species:Mus musculus	RAW_FILE_NAME=221125_AceCoA_diff_3T3L1_Alix_Set2.wiff
SUBJECT_SAMPLE_FACTORS           	-	3T3L1_ctrl_d10_4	Treatment:d1 | Replicate:4 | Species:Mus musculus	RAW_FILE_NAME=221125_AceCoA_diff_3T3L1_Alix_Set2.wiff
SUBJECT_SAMPLE_FACTORS           	-	3T3L1_ctrl_d10_5	Treatment:d1 | Replicate:5 | Species:Mus musculus	RAW_FILE_NAME=221123_AceCoA_diff_3T3L1_Alix.wiff
SUBJECT_SAMPLE_FACTORS           	-	3T3L1_ctrl_d10_6	Treatment:d1 | Replicate:6 | Species:Mus musculus	RAW_FILE_NAME=221123_AceCoA_diff_3T3L1_Alix.wiff
#COLLECTION
CO:COLLECTION_SUMMARY            	Human SGBS cells were provided by the laboratory of Prof. Dr. Wabitsch at the
CO:COLLECTION_SUMMARY            	University Clinic Ulm. Cells were differentiated according to the standard
CO:COLLECTION_SUMMARY            	protocol described previously (Wabitsch et al., 2001). 3T3-L1 mouse embryonic
CO:COLLECTION_SUMMARY            	fibroblasts were purchased from the American Type Culture Collection (ATCC,
CO:COLLECTION_SUMMARY            	CL-173, USA).
CO:SAMPLE_TYPE                   	Adipose tissue
CO:STORAGE_CONDITIONS            	-80℃
#TREATMENT
TR:TREATMENT_SUMMARY             	Both cell lines were maintained under 5% CO2 at 37°C and 95% humidity.
TR:TREATMENT_SUMMARY             	Cell culture experiments for metabolomic (SGBS and 3T3L1) data analysis were
TR:TREATMENT_SUMMARY             	performed in 6 replicates. Cells were cultivated until day0,day6 and day12 for
TR:TREATMENT_SUMMARY             	the SGBS cell line, and until day0, day5 and day10 for the 3T3-L1 cells.
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	Intracellular metabolites were extracted using a 1:1:1 methanol:water:chloroform
SP:SAMPLEPREP_SUMMARY            	extraction protocol. Briefly, culture media was removed and cells were rinsed
SP:SAMPLEPREP_SUMMARY            	with icecold 0.9 % sodium chloride solution. The rinsing solution was removed
SP:SAMPLEPREP_SUMMARY            	and the cells metabolism was quenched by adding equal volumes of methanol,
SP:SAMPLEPREP_SUMMARY            	followed by icecold deionized water. Cells were scraped off the culture plate
SP:SAMPLEPREP_SUMMARY            	and combined chloroform. Samples were vortexed and kept on a shaker at 4°C for
SP:SAMPLEPREP_SUMMARY            	20 min at 1400 rpm. Cell debris was removed by centrifugation. A fixed
SP:SAMPLEPREP_SUMMARY            	volume of the polar upper phase was transferred to new tubes and dried to
SP:SAMPLEPREP_SUMMARY            	completeness.
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_TYPE           	Reversed phase
CH:INSTRUMENT_NAME               	Agilent 1290 Infinity II
CH:COLUMN_NAME                   	Waters XSelect XP HSS T3 (150 x 2.1mm, 2.5um)
CH:SOLVENT_A                     	10mM TBA, 10mM acetic acid, 5% MeOH, 2% IPA in water
CH:SOLVENT_B                     	100% IPA
CH:FLOW_GRADIENT                 	0-5 min 0% B, 5-9 min 0%- 2% B, 9-9.5 min 2-6% B, 9.5-11.5 min 6% B, 11.5-12 min
CH:FLOW_GRADIENT                 	6-11% B, 12-13.5 min 11% B, 13.5-15.5 min 11-28% B, 15.5-16.5 min 28-53% B,
CH:FLOW_GRADIENT                 	16.5-22.5 53% B, 22.5-23 min 53-0% B, 23-33 min 0% B
CH:FLOW_RATE                     	0-15.5 min 0.4 mL/min, 15.5-16.5 min 0.4-0.15 mL/min, 16.5-23 min 0.15 mL/min,
CH:FLOW_RATE                     	23-27 min 0.15-0.4 mL/min, 27-33 min 0.4 mL/min
CH:COLUMN_TEMPERATURE            	40
#ANALYSIS
AN:ANALYSIS_TYPE                 	MS
#MS
MS:INSTRUMENT_NAME               	ABI Sciex 6500+
MS:INSTRUMENT_TYPE               	Triple quadrupole
MS:MS_TYPE                       	ESI
MS:ION_MODE                      	NEGATIVE
MS:MS_COMMENTS                   	For identification and quantitation, a scheduled MRM method was used, with
MS:MS_COMMENTS                   	specific transitions for every metabolite. Data acquisition and peak integration
MS:MS_COMMENTS                   	were performed in Analyst® software (Version 1.7.1).
#MS_METABOLITE_DATA
MS_METABOLITE_DATA:UNITS	AUC
MS_METABOLITE_DATA_START
Samples	SGBS_ctrl_d0_1	SGBS_ctrl_d0_2	SGBS_ctrl_d0_3	SGBS_ctrl_d0_4	SGBS_ctrl_d0_5	SGBS_ctrl_d0_6	SGBS_ctrl_d3_1	SGBS_ctrl_d3_2	SGBS_ctrl_d3_3	SGBS_ctrl_d3_4	SGBS_ctrl_d3_5	SGBS_ctrl_d3_6	SGBS_ctrl_d6_1	SGBS_ctrl_d6_2	SGBS_ctrl_d6_3	SGBS_ctrl_d6_4	SGBS_ctrl_d6_5	SGBS_ctrl_d6_6	SGBS_ctrl_d9_1	SGBS_ctrl_d9_2	SGBS_ctrl_d9_3	SGBS_ctrl_d9_4	SGBS_ctrl_d9_5	SGBS_ctrl_d9_6	SGBS_ctrl_d12_1	SGBS_ctrl_d12_2	SGBS_ctrl_d12_3	SGBS_ctrl_d12_4	SGBS_ctrl_d12_5	SGBS_ctrl_d12_6	3T3L1_ctrl_d0_1	3T3L1_ctrl_d0_2	3T3L1_ctrl_d0_3	3T3L1_ctrl_d0_4	3T3L1_ctrl_d0_5	3T3L1_ctrl_d0_6	3T3L1_ctrl_d3_1	3T3L1_ctrl_d3_2	3T3L1_ctrl_d3_3	3T3L1_ctrl_d3_4	3T3L1_ctrl_d3_5	3T3L1_ctrl_d3_6	3T3L1_ctrl_d5_1	3T3L1_ctrl_d5_2	3T3L1_ctrl_d5_3	3T3L1_ctrl_d5_4	3T3L1_ctrl_d5_5	3T3L1_ctrl_d5_6	3T3L1_ctrl_d7_1	3T3L1_ctrl_d7_2	3T3L1_ctrl_d7_3	3T3L1_ctrl_d7_4	3T3L1_ctrl_d7_5	3T3L1_ctrl_d7_6	3T3L1_ctrl_d10_1	3T3L1_ctrl_d10_2	3T3L1_ctrl_d10_3	3T3L1_ctrl_d10_4	3T3L1_ctrl_d10_5	3T3L1_ctrl_d10_6
Factors	Treatment:d0 | Replicate:1 | Species:Homo sapiens	Treatment:d0 | Replicate:2 | Species:Homo sapiens	Treatment:d0 | Replicate:3 | Species:Homo sapiens	Treatment:d0 | Replicate:4 | Species:Homo sapiens	Treatment:d0 | Replicate:5 | Species:Homo sapiens	Treatment:d0 | Replicate:6 | Species:Homo sapiens	Treatment:d3 | Replicate:1 | Species:Homo sapiens	Treatment:d3 | Replicate:2 | Species:Homo sapiens	Treatment:d3 | Replicate:3 | Species:Homo sapiens	Treatment:d3 | Replicate:4 | Species:Homo sapiens	Treatment:d3 | Replicate:5 | Species:Homo sapiens	Treatment:d3 | Replicate:6 | Species:Homo sapiens	Treatment:d6 | Replicate:1 | Species:Homo sapiens	Treatment:d6 | Replicate:2 | Species:Homo sapiens	Treatment:d6 | Replicate:3 | Species:Homo sapiens	Treatment:d6 | Replicate:4 | Species:Homo sapiens	Treatment:d6 | Replicate:5 | Species:Homo sapiens	Treatment:d6 | Replicate:6 | Species:Homo sapiens	Treatment:d9 | Replicate:1 | Species:Homo sapiens	Treatment:d9 | Replicate:2 | Species:Homo sapiens	Treatment:d9 | Replicate:3 | Species:Homo sapiens	Treatment:d9 | Replicate:4 | Species:Homo sapiens	Treatment:d9 | Replicate:5 | Species:Homo sapiens	Treatment:d9 | Replicate:6 | Species:Homo sapiens	Treatment:d12 | Replicate:1 | Species:Homo sapiens	Treatment:d12 | Replicate:2 | Species:Homo sapiens	Treatment:d12 | Replicate:3 | Species:Homo sapiens	Treatment:d12 | Replicate:4 | Species:Homo sapiens	Treatment:d12 | Replicate:5 | Species:Homo sapiens	Treatment:d12 | Replicate:6 | Species:Homo sapiens	Treatment:d0 | Replicate:1 | Species:Mus musculus	Treatment:d0 | Replicate:2 | Species:Mus musculus	Treatment:d0 | Replicate:3 | Species:Mus musculus	Treatment:d0 | Replicate:4 | Species:Mus musculus	Treatment:d0 | Replicate:5 | Species:Mus musculus	Treatment:d0 | Replicate:6 | Species:Mus musculus	Treatment:d3 | Replicate:1 | Species:Mus musculus	Treatment:d3 | Replicate:2 | Species:Mus musculus	Treatment:d3 | Replicate:3 | Species:Mus musculus	Treatment:d3 | Replicate:4 | Species:Mus musculus	Treatment:d3 | Replicate:5 | Species:Mus musculus	Treatment:d3 | Replicate:6 | Species:Mus musculus	Treatment:d5 | Replicate:1 | Species:Mus musculus	Treatment:d5 | Replicate:2 | Species:Mus musculus	Treatment:d5 | Replicate:3 | Species:Mus musculus	Treatment:d5 | Replicate:4 | Species:Mus musculus	Treatment:d5 | Replicate:5 | Species:Mus musculus	Treatment:d5 | Replicate:6 | Species:Mus musculus	Treatment:d7 | Replicate:1 | Species:Mus musculus	Treatment:d7 | Replicate:2 | Species:Mus musculus	Treatment:d7 | Replicate:3 | Species:Mus musculus	Treatment:d7 | Replicate:4 | Species:Mus musculus	Treatment:d7 | Replicate:5 | Species:Mus musculus	Treatment:d7 | Replicate:6 | Species:Mus musculus	Treatment:d1 | Replicate:1 | Species:Mus musculus	Treatment:d1 | Replicate:2 | Species:Mus musculus	Treatment:d1 | Replicate:3 | Species:Mus musculus	Treatment:d1 | Replicate:4 | Species:Mus musculus	Treatment:d1 | Replicate:5 | Species:Mus musculus	Treatment:d1 | Replicate:6 | Species:Mus musculus
Glucose-6-phosphate	1.39E+06	1.65E+06	1.56E+06	1.49E+06	1.55E+06	1.41E+06	1.68E+05	1.73E+05	4.23E+05	1.72E+05	2.26E+05	1.80E+05	5.32E+05	5.37E+05	5.24E+05	5.56E+05	5.28E+05	5.62E+05	1.77E+06	1.42E+06	1.82E+06	1.55E+06	1.77E+06	2.49E+06	9.89E+06	1.09E+07	1.12E+07		1.01E+07	9.75E+06	1.86E+06	2.06E+06	2.21E+06	1.65E+06	2.02E+06	2.05E+06	5.78E+06	7.50E+06	6.48E+06	6.61E+06	6.63E+06	6.07E+06	2.16E+07	1.87E+07	1.99E+07	1.96E+07	2.02E+07	2.05E+07	2.68E+07	2.48E+07	2.51E+07	2.75E+07	2.60E+07	2.88E+07		2.48E+07	2.79E+07	2.41E+07	2.99E+07	2.84E+07
Citrate	3.87E+07	3.57E+07	4.22E+07	4.14E+07	4.42E+07	4.18E+07	1.91E+07	2.19E+07	6.03E+07	1.63E+07	1.88E+07	2.29E+07	7.64E+06	1.09E+07	7.91E+06	8.63E+06	7.55E+06	9.14E+06	1.09E+07	8.61E+06	1.39E+07	7.31E+06	9.20E+06	1.15E+07	4.56E+06	4.95E+06	5.00E+06	4.75E+06	5.62E+06	6.38E+06	3.76E+07	4.26E+07	3.93E+07	3.31E+07	3.69E+07	3.84E+07	3.42E+08	3.74E+08	3.39E+08	3.41E+08	3.58E+08	3.10E+08	5.40E+08	4.88E+08	5.03E+08	4.90E+08	5.20E+08	5.22E+08	5.48E+08	5.42E+08	5.29E+08	5.41E+08	5.56E+08	6.13E+08	4.86E+08	4.83E+08	4.81E+08	4.64E+08	4.43E+08	5.09E+08
a-Ketoglutarate	2.84E+06	3.16E+06	3.57E+06	3.60E+06	3.44E+06	3.60E+06	5.53E+05	7.31E+05	7.02E+05	6.81E+05	7.06E+05	6.72E+05	1.29E+06	1.39E+06	1.42E+06	1.51E+06	1.39E+06	1.59E+06	4.95E+05	4.56E+05	4.86E+05	4.70E+05	4.97E+05	5.02E+05	4.12E+05	4.55E+05	4.21E+05	6.70E+05	4.50E+05	5.41E+05	3.53E+06	3.66E+06	3.67E+06	3.35E+06	3.36E+06	4.21E+06	2.16E+07	2.27E+07	2.01E+07	2.24E+07	1.94E+07	1.73E+07	3.89E+07	2.93E+07	3.62E+07	3.34E+07	2.89E+07	3.05E+07	3.09E+07	3.32E+07	3.23E+07	4.00E+07	3.22E+07	4.37E+07	1.62E+07	2.08E+07	1.57E+07	2.24E+07	1.63E+07	2.41E+07
Succinate	5.13E+06	4.32E+06	4.84E+06	4.56E+06	7.26E+06	4.29E+06	8.24E+05	7.70E+05	9.12E+05	8.39E+05	7.91E+05	7.38E+05	1.10E+06	1.30E+06	1.09E+06	1.06E+06	1.09E+06	1.11E+06	5.72E+05	7.18E+05	7.74E+05	6.21E+05	9.17E+05	7.24E+05	6.02E+05	5.74E+05	1.06E+06	6.73E+05	4.88E+05	5.58E+05	1.04E+07	1.41E+07	2.22E+07	9.19E+06	1.10E+07	1.42E+07	4.41E+07	5.65E+07	5.09E+07	4.51E+07	4.46E+07	4.76E+07	7.24E+07	6.08E+07	6.56E+07	6.77E+07	6.52E+07	6.68E+07	1.09E+08	1.09E+08	9.99E+07	1.15E+08	1.11E+08	1.19E+08	2.55E+08	2.24E+08	2.56E+08	2.30E+08	2.81E+08	2.48E+08
Phosphoenolpyruvate	4.47E+05	5.41E+05	5.11E+05	5.51E+05	5.25E+05	5.46E+05	1.84E+05	1.98E+05	2.01E+05	2.06E+05	1.96E+05	2.15E+05	8.28E+05	9.84E+05	8.68E+05	1.00E+06	8.07E+05	8.81E+05	1.88E+06	1.65E+06	1.81E+06	1.54E+06	1.51E+06	1.98E+06	1.11E+06	1.60E+06	1.38E+06	4.89E+05	1.49E+06	1.32E+06	7.04E+05	7.47E+05	7.48E+05	7.31E+05	6.70E+05	7.92E+05	2.47E+06	2.60E+06	1.91E+06	1.87E+06	2.24E+06	1.64E+06	4.54E+06	3.17E+06	5.09E+06	5.12E+06	5.23E+06	6.11E+06	3.00E+06	5.08E+06	2.16E+06	3.30E+06	3.38E+06	4.07E+06	4.52E+06	2.29E+06	2.34E+06	1.24E+06	2.28E+06	2.52E+06
Pyruvate	2.16E+05	2.37E+05	2.41E+05	2.42E+05	2.70E+05	2.31E+05	5.51E+05	7.73E+05	2.06E+06	5.57E+05	6.42E+05	7.80E+05	1.82E+05	1.67E+05	1.82E+05	1.97E+05	2.16E+05	2.46E+05	3.98E+05	3.79E+05	4.41E+05	3.52E+05	3.80E+05	4.38E+05	5.22E+05	5.40E+05	5.43E+05	2.03E+05	5.31E+05	4.92E+05	1.16E+06	1.02E+06	1.80E+06	1.03E+06	1.09E+06	1.61E+06	1.35E+06	1.93E+06	1.20E+06	1.98E+06	1.17E+06	1.66E+06	7.55E+06	4.72E+06	5.40E+06	6.71E+06	5.45E+06	6.45E+06	2.58E+06	3.98E+06	3.38E+06	4.74E+06	3.74E+06	5.86E+06	1.53E+06	1.88E+06	1.43E+06	1.98E+06	1.62E+06	2.50E+06
Fructose-6-phosphate	1.99E+05	2.12E+05	1.87E+05	2.44E+05	2.87E+05	2.12E+05	7.16E+03	1.16E+04	3.77E+04	7.23E+03	1.34E+04	8.43E+03	9.51E+04	8.72E+04	7.90E+04	6.80E+04	7.16E+04	9.08E+04	3.77E+05	1.74E+05	3.78E+05	2.65E+05	3.19E+05	5.18E+05	1.91E+06	2.18E+06	2.29E+06	1.49E+05	1.97E+06	1.94E+06	6.29E+05	6.48E+05	6.31E+05	4.01E+05	6.38E+05	6.96E+05	2.42E+06	3.51E+06	3.08E+06	2.42E+06	2.66E+06	2.12E+06	7.31E+06	7.71E+06	1.02E+07	8.01E+06	8.49E+06	7.18E+06	6.78E+06	7.14E+06	8.81E+06	8.91E+06	9.91E+06	7.77E+06	1.07E+07	7.65E+06	9.02E+06	5.89E+06	7.92E+06	7.95E+06
Glyceraldehyde-3-phosphate	3.62E+04	3.65E+04	4.59E+04	5.18E+04	5.42E+04	7.12E+03	8.16E+03	8.99E+03	1.99E+04	6.94E+03	8.95E+03	7.37E+03	1.01E+05	1.05E+05	9.83E+04	1.09E+05	7.91E+04	1.07E+05	1.46E+05	2.11E+05	1.66E+05	1.58E+05	1.57E+05	1.84E+05	2.40E+05	4.00E+05	2.55E+05	9.63E+04	3.30E+05	2.97E+05	3.11E+04	3.15E+04	2.19E+04	2.12E+04	2.75E+04	3.53E+04	2.86E+05	3.45E+05	3.08E+05	3.40E+05	3.04E+05	3.04E+05	6.59E+05	5.09E+05	5.35E+05	6.15E+05	5.27E+05	5.89E+05	3.16E+05	3.50E+05	3.58E+05	4.66E+05	3.34E+05	4.58E+05	8.77E+05	3.05E+05	2.79E+05	3.36E+05	2.14E+05	2.58E+05
Fumarate	2.35E+05	2.71E+05	2.64E+05	2.45E+05	2.38E+05	2.36E+05	9.95E+04	1.06E+05	1.22E+05	1.04E+05	1.09E+05	1.04E+05	7.10E+04	7.76E+04	7.23E+04	7.67E+04	7.23E+04	7.60E+04	1.10E+05	1.06E+05	1.05E+05	1.04E+05	9.80E+04	1.13E+05	8.23E+05	1.29E+06	1.05E+06	1.07E+05	9.56E+05	9.63E+05	2.38E+05	2.29E+05	1.89E+05	2.03E+05	2.24E+05	2.72E+05	3.16E+06	3.43E+06	3.03E+06	3.14E+06	3.16E+06	2.84E+06	5.63E+06	5.00E+06	5.32E+06	5.18E+06	5.18E+06	5.63E+06	5.25E+06	4.83E+06	4.89E+06	5.74E+06	5.28E+06	5.67E+06	2.75E+06	2.46E+06	2.73E+06	2.63E+06	2.99E+06	2.74E+06
Oxaloacetate	5.68E+04	5.88E+04	5.88E+04	6.49E+04	5.57E+04	5.38E+04	4.77E+04	2.90E+04	2.38E+04	2.30E+04	1.81E+04	1.88E+04	1.80E+05	1.73E+05	1.87E+05	1.88E+05	1.69E+05	1.82E+05	1.56E+05	1.47E+05	1.73E+05	1.46E+05	1.39E+05	1.68E+05	1.41E+05	1.71E+05	1.56E+05	7.51E+04	1.55E+05	1.51E+05	1.04E+05	5.88E+04	1.05E+05	7.03E+04	8.06E+04	2.11E+05	1.30E+05	3.17E+05	2.70E+05	2.64E+05	3.09E+05	2.48E+05	5.70E+05	5.76E+05	3.52E+05	5.46E+05	5.95E+05	4.81E+05	7.31E+05	6.46E+05	3.74E+05	5.60E+05	6.54E+05	3.51E+05	2.74E+05	3.08E+05	3.83E+05	2.07E+05	4.87E+05	3.78E+05
Fructose-1,6-bisphosphate	1.31E+07	1.49E+07	1.70E+07	1.56E+07	1.53E+07	1.48E+07	2.58E+05	3.90E+05	6.22E+05	3.34E+05	6.16E+05	2.17E+05	1.01E+06	1.17E+06	1.09E+06	8.04E+05	9.01E+05	8.94E+05	4.23E+06	2.68E+06	3.84E+06	3.15E+06	3.54E+06	6.15E+06	1.29E+07	1.22E+07	1.30E+07	1.32E+06	1.16E+07	1.33E+07	2.06E+07	2.02E+07	1.92E+07	1.92E+07	1.72E+07	1.96E+07	7.67E+07	5.82E+07	6.45E+07	6.86E+07	7.25E+07	5.82E+07	1.51E+08	1.40E+08	1.43E+08	1.31E+08	1.25E+08	1.32E+08	1.09E+08	8.16E+07	9.86E+07	1.07E+08	8.19E+07	1.14E+08	4.84E+06	5.17E+07	6.01E+07	7.60E+07	5.27E+07	8.69E+07
Cis-Aconitate	2.40E+06	2.43E+06	2.51E+06	2.42E+06	2.44E+06	2.40E+06	2.28E+05	2.40E+05	3.26E+05	2.35E+05	2.95E+05	2.44E+05	4.22E+05	4.85E+05	4.41E+05	4.84E+05	4.52E+05	4.82E+05	5.73E+05	7.23E+05	8.62E+05	4.50E+05	6.06E+05	6.11E+05	3.41E+05	4.81E+05	3.79E+05	3.44E+05	3.78E+05	4.11E+05	1.39E+06	1.54E+06	1.61E+06	1.30E+06	1.33E+06	1.68E+06	1.43E+07	1.57E+07	1.38E+07	1.39E+07	1.45E+07	1.25E+07	2.51E+07	2.08E+07	2.39E+07	2.38E+07	2.35E+07	2.35E+07	2.73E+07	2.65E+07	2.65E+07	2.85E+07	2.91E+07	3.34E+07	2.19E+07	2.04E+07	2.02E+07	1.90E+07	1.93E+07	2.29E+07
Malate	4.28E+07	4.49E+07	4.56E+07	4.38E+07	4.62E+07	4.24E+07	1.69E+07	1.76E+07	2.01E+07	1.79E+07	1.75E+07	1.76E+07	1.37E+07	1.58E+07	1.54E+07	1.57E+07	1.38E+07	1.51E+07	2.23E+07	1.83E+07	2.43E+07	1.92E+07	1.75E+07	2.32E+07	1.22E+08	1.68E+08	1.44E+08	2.00E+07	1.48E+08	1.42E+08	5.68E+07	5.95E+07	4.63E+07	5.02E+07	5.56E+07	6.02E+07	4.09E+08	4.50E+08	4.07E+08	4.20E+08	4.16E+08	3.90E+08	6.12E+08	5.55E+08	5.78E+08	5.73E+08	5.81E+08	5.84E+08	5.34E+08	5.35E+08	5.33E+08	5.80E+08	5.66E+08	5.82E+08	4.08E+08	3.89E+08	4.09E+08	3.81E+08	4.15E+08	4.03E+08
Acetyl-CoA	2.31E+05	2.46E+05	2.26E+05	1.99E+05	2.42E+05	1.87E+05	2.37E+05	2.51E+05	2.79E+05	2.51E+05	2.71E+05	2.42E+05	9.98E+05	4.07E+05	8.37E+05	5.12E+05	8.29E+05	7.90E+05	1.34E+06	1.37E+06	1.25E+06	1.50E+06	1.36E+06	1.57E+06	1.37E+06	1.57E+06	1.46E+06	6.10E+05	1.50E+06	1.38E+06	4.85E+05	4.22E+05	4.77E+05	4.94E+05	4.57E+05	5.75E+05	5.37E+06	3.08E+06	4.36E+06	3.87E+06	4.68E+06	3.91E+06	9.26E+06	7.75E+06	1.06E+07	7.55E+06	7.57E+06	6.33E+06	5.73E+06	3.99E+06	5.64E+06	6.03E+06	4.95E+06	5.95E+06	1.57E+06	1.34E+06	1.28E+06	2.39E+06	1.31E+06	1.70E+06
3-Phosphoglycerate	3.63E+05	4.67E+05	4.45E+05	5.15E+05	5.51E+05	4.87E+05	1.63E+05	1.67E+05	2.06E+05	2.13E+05	1.78E+05	1.98E+05	8.02E+05	8.51E+05	8.10E+05	9.61E+05	8.05E+05	1.01E+06	2.13E+06	1.45E+06	2.36E+06	1.67E+06	1.62E+06	2.56E+06	1.07E+06	1.35E+06	1.28E+06	4.55E+05	1.39E+06	1.30E+06	1.21E+06	1.26E+06	1.25E+06	1.23E+06	1.16E+06	1.34E+06	4.03E+06	4.55E+06	3.38E+06	3.31E+06	4.03E+06	2.97E+06	7.93E+06	5.70E+06	8.77E+06	8.68E+06	8.69E+06	1.07E+07	5.07E+06	8.75E+06	3.53E+06	5.59E+06	5.46E+06	6.62E+06	7.79E+06	3.68E+06	3.65E+06	2.05E+06	3.59E+06	4.04E+06
MS_METABOLITE_DATA_END
#METABOLITES
METABOLITES_START
metabolite_name	m/z	KEGG_ID
Glucose-6-phosphate	259	C00668 
Citrate	191	C00158
a-Ketoglutarate	145	C00026 
Succinate	117	C00042
Phosphoenolpyruvate	167	C00074
Pyruvate	87	C00022
Fructose-6-phosphate	259	C00085 
Glyceraldehyde-3-phosphate	169	C00118   
Fumarate	115	C00122
Oxaloacetate	131	C00036
Fructose-1,6-bisphosphate	339	C05378 
Cis-Aconitate	173	C00417 
Malate	133	C00711
Acetyl-CoA	808	C00024
3-Phosphoglycerate	185	C00197 
METABOLITES_END
#END