#METABOLOMICS WORKBENCH engelmab_20250424_021839 DATATRACK_ID:5860 STUDY_ID:ST003879 ANALYSIS_ID:AN006372 PROJECT_ID:PR002433
VERSION             	1
CREATED_ON             	April 28, 2025, 6:45 pm
#PROJECT
PR:PROJECT_TITLE                 	PLCXD1 as a Regulator of Insulin-Stimulated Lipid Metabolism in Human Adipocytes
PR:PROJECT_SUMMARY               	Insulin regulates adipocyte biology through transcriptional and
PR:PROJECT_SUMMARY               	post-transcriptional mechanisms, yet many insulin-responsive targets remain
PR:PROJECT_SUMMARY               	poorly characterized. Phospholipase C X domain-containing protein-1 (PLCXD1)
PR:PROJECT_SUMMARY               	emerged as a regulator of insulin-stimulated lipogenesis, without affecting
PR:PROJECT_SUMMARY               	lipolysis or adipogenesis. Our findings establish PLCXD1 as an
PR:PROJECT_SUMMARY               	insulin-responsive effector of lipid metabolism, with implications for adipocyte
PR:PROJECT_SUMMARY               	dysfunction in insulin resistance.
PR:INSTITUTE                     	Helmholtz Centre for Environmental Research
PR:LAST_NAME                     	Engelmann
PR:FIRST_NAME                    	Beatrice
PR:ADDRESS                       	Permoserstr. 15
PR:EMAIL                         	beatrice.engelmann@ufz.de
PR:PHONE                         	00493412351099
#STUDY
ST:STUDY_TITLE                   	Establishing an insulin-sensitive human adipocyte culture model
ST:STUDY_SUMMARY                 	To enable functional screens of insulin action in human fat cells, we generated
ST:STUDY_SUMMARY                 	adipocytes from adipose progenitor cells in vitro. Under standardized culture
ST:STUDY_SUMMARY                 	conditions, the adipocytes displayed limited insulin responsiveness. Our results
ST:STUDY_SUMMARY                 	prompted us to establish optimal insulin sensitizing conditions. One of the
ST:STUDY_SUMMARY                 	tested conditions resulted in fat cells with enhanced insulin responsiveness. To
ST:STUDY_SUMMARY                 	confirm that our results reflected expected effects in energy metabolism, we
ST:STUDY_SUMMARY                 	traced glucose conversion into central carbon metabolites following two hours of
ST:STUDY_SUMMARY                 	insulin stimulation. This revealed a robust increase in labeling of glycolytic
ST:STUDY_SUMMARY                 	and tricarboxylic acid (TCA) cycle intermediates in insulin-treated versus
ST:STUDY_SUMMARY                 	untreated cells.
ST:INSTITUTE                     	Helmholtz Centre for Environmental Research
ST:LAST_NAME                     	Engelmann
ST:FIRST_NAME                    	Beatrice
ST:ADDRESS                       	Permoserstr. 15
ST:EMAIL                         	beatrice.engelmann@ufz.de
ST:PHONE                         	00493412351099
#SUBJECT
SU:SUBJECT_TYPE                  	Cultured cells
SU:SUBJECT_SPECIES               	Homo sapiens
SU:TAXONOMY_ID                   	9606
SU:GENDER                        	Male
#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           	-	2_Insulin_sgCON	Tretament:Insulin | Sample source:subcutaneous WAT	Replicate=1; Species=Homo sapiens; RAW_FILE_NAME=231006_13C_MIDAS_13C-glucose_cells.wiff
SUBJECT_SAMPLE_FACTORS           	-	9_Insulin_sgCON	Tretament:Insulin | Sample source:subcutaneous WAT	Replicate=2; Species=Homo sapiens; RAW_FILE_NAME=231006_13C_MIDAS_13C-glucose_cells.wiff
SUBJECT_SAMPLE_FACTORS           	-	5_Insulin_sgCON	Tretament:Insulin | Sample source:subcutaneous WAT	Replicate=3; Species=Homo sapiens; RAW_FILE_NAME=231006_13C_MIDAS_13C-glucose_cells.wiff
SUBJECT_SAMPLE_FACTORS           	-	10_Insulin_sgCON	Tretament:Insulin | Sample source:subcutaneous WAT	Replicate=4; Species=Homo sapiens; RAW_FILE_NAME=231006_13C_MIDAS_13C-glucose_cells.wiff
SUBJECT_SAMPLE_FACTORS           	-	1_Insulin_sgCON	Tretament:Insulin | Sample source:subcutaneous WAT	Replicate=5; Species=Homo sapiens; RAW_FILE_NAME=231006_13C_MIDAS_13C-glucose_cells.wiff
SUBJECT_SAMPLE_FACTORS           	-	6_Insulin_sgCON	Tretament:Insulin | Sample source:subcutaneous WAT	Replicate=6; Species=Homo sapiens; RAW_FILE_NAME=231006_13C_MIDAS_13C-glucose_cells.wiff
SUBJECT_SAMPLE_FACTORS           	-	21_Basal_sgCON	Tretament:Basal | Sample source:subcutaneous WAT	Replicate=1; Species=Homo sapiens; RAW_FILE_NAME=231006_13C_MIDAS_13C-glucose_cells.wiff
SUBJECT_SAMPLE_FACTORS           	-	13_Basal_sgCON	Tretament:Basal | Sample source:subcutaneous WAT	Replicate=2; Species=Homo sapiens; RAW_FILE_NAME=231006_13C_MIDAS_13C-glucose_cells.wiff
SUBJECT_SAMPLE_FACTORS           	-	14_Basal_sgCON	Tretament:Basal | Sample source:subcutaneous WAT	Replicate=3; Species=Homo sapiens; RAW_FILE_NAME=231006_13C_MIDAS_13C-glucose_cells.wiff
SUBJECT_SAMPLE_FACTORS           	-	17_Basal_sgCON	Tretament:Basal | Sample source:subcutaneous WAT	Replicate=4; Species=Homo sapiens; RAW_FILE_NAME=231006_13C_MIDAS_13C-glucose_cells.wiff
SUBJECT_SAMPLE_FACTORS           	-	18_Basal_sgCON	Tretament:Basal | Sample source:subcutaneous WAT	Replicate=5; Species=Homo sapiens; RAW_FILE_NAME=231006_13C_MIDAS_13C-glucose_cells.wiff
SUBJECT_SAMPLE_FACTORS           	-	22_Basal_sgCON	Tretament:Basal | Sample source:subcutaneous WAT	Replicate=6; Species=Homo sapiens; RAW_FILE_NAME=231006_13C_MIDAS_13C-glucose_cells.wiff
#COLLECTION
CO:COLLECTION_SUMMARY            	Human preadipocytes were obtained from abdominal subcutaneous white adipose
CO:COLLECTION_SUMMARY            	tissue (WAT) of a male patient.
CO:SAMPLE_TYPE                   	Subcutaneous white adipose tissue
#TREATMENT
TR:TREATMENT_SUMMARY             	Cells were cultured in 5% CO2 at 37°C. Cells were proliferated in DMEM with 10
TR:TREATMENT_SUMMARY             	mmol/l HEPES, 10% FBS, 50 μg/mL penicillin/streptomycin and 2.5 ng/mL
TR:TREATMENT_SUMMARY             	Fibroblast growth factor-2 (FGF2). FGF was removed 24h prior to initiating
TR:TREATMENT_SUMMARY             	differentiation. To initiate differentiation, media was switched to a 50/50
TR:TREATMENT_SUMMARY             	mixture of DMEM/F12 with 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid
TR:TREATMENT_SUMMARY             	(HEPES) and insulin, triiodothyronine (T3), transferrin, dexamethasone,
TR:TREATMENT_SUMMARY             	3-isobutyl-1-methylxanthine (IBMX) and rosiglitazone for three days, after which
TR:TREATMENT_SUMMARY             	dexamethasone and IBMX were removed. Rosiglitazone was removed on day 10 of
TR:TREATMENT_SUMMARY             	differentiation. Various T3, transferrin and insulin supplementation conditions
TR:TREATMENT_SUMMARY             	were compared for insulin sensitization, so that afterwards all experiments were
TR:TREATMENT_SUMMARY             	performed with T3 and transferrin maintained in the media throughout
TR:TREATMENT_SUMMARY             	differentiation, while insulin was reduced on day 10 of differentiation. On D13,
TR:TREATMENT_SUMMARY             	cells were washed once with DMEM no glucose/glutamine supplemented with HEPES
TR:TREATMENT_SUMMARY             	and PEST prior to incubation in DMEM with glucose and for 3h at 37°C. Cells
TR:TREATMENT_SUMMARY             	were washed with DMEM no glucose/no glutamine and media replaced with tracing
TR:TREATMENT_SUMMARY             	media containing labeled C13-glucose and unlabeled glutamine with or without
TR:TREATMENT_SUMMARY             	insulin.
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	Extraction of intracellular and extracellular metabolites was performed by a
SP:SAMPLEPREP_SUMMARY            	1:1:1 methanol:water:chloroform extraction protocol. For the extraction of
SP:SAMPLEPREP_SUMMARY            	intracellular metabolites, the culture medium was removed and the cells were
SP:SAMPLEPREP_SUMMARY            	rinsed twice with 1 ml of 0.9% ice-cold NaCl. The rinsing solution was removed,
SP:SAMPLEPREP_SUMMARY            	and the metabolism of the cells was stopped by adding 400 µL of MeOH (-20 °C)
SP:SAMPLEPREP_SUMMARY            	followed by 400 µL of ice-cold H2O containing 10 µM d6-glutarate. Cells were
SP:SAMPLEPREP_SUMMARY            	collected using a cell lifter and 400 µL of chloroform was added. After shaking
SP:SAMPLEPREP_SUMMARY            	at 1,400 rpm and 4 °C for 20 min, the extraction mixture was centrifuged at
SP:SAMPLEPREP_SUMMARY            	18,000 g and 4 °C for 5 min. Subsequently, the polar upper phase was collected
SP:SAMPLEPREP_SUMMARY            	and evaporated to complete dryness.
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_SUMMARY        	Solvent A: 10mM tributylamine, 10mM acetic acid, 5% MeOH, 2% 2-propanol in
CH:CHROMATOGRAPHY_SUMMARY        	water; Solvent B: 100% 2-propanol
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                     	93% water/5% methanol/2% isopropanol; 10mM tributylamine; 10mM acetic acid
CH:SOLVENT_B                     	100% isopropanol
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               	QTRAP
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 isotopologue and metabolite. Every
MS:MS_COMMENTS                   	precursor-fragment pair is defined by the amount of labeled carbons in the
MS:MS_COMMENTS                   	parent ion (Mx) and in the fragment ion (Fx). Data acquisition was performed in
MS:MS_COMMENTS                   	Analyst® software (Version 1.7.1) and Data integration in SciexOS(R) (Version
MS:MS_COMMENTS                   	3.0.0).
#MS_METABOLITE_DATA
MS_METABOLITE_DATA:UNITS	Peak Area
MS_METABOLITE_DATA_START
Samples	1_Insulin_sgCON	10_Insulin_sgCON	2_Insulin_sgCON	5_Insulin_sgCON	6_Insulin_sgCON	9_Insulin_sgCON	13_Basal_sgCON	14_Basal_sgCON	17_Basal_sgCON	18_Basal_sgCON	21_Basal_sgCON	22_Basal_sgCON
Factors	Tretament:Insulin | Sample source:subcutaneous WAT	Tretament:Insulin | Sample source:subcutaneous WAT	Tretament:Insulin | Sample source:subcutaneous WAT	Tretament:Insulin | Sample source:subcutaneous WAT	Tretament:Insulin | Sample source:subcutaneous WAT	Tretament:Insulin | Sample source:subcutaneous WAT	Tretament:Basal | Sample source:subcutaneous WAT	Tretament:Basal | Sample source:subcutaneous WAT	Tretament:Basal | Sample source:subcutaneous WAT	Tretament:Basal | Sample source:subcutaneous WAT	Tretament:Basal | Sample source:subcutaneous WAT	Tretament:Basal | Sample source:subcutaneous WAT
Citrate_M0	1.17E+06	1.34E+06	1.27E+06	1.12E+06	1.15E+06	1.21E+06	1.19E+06	1.23E+06	1.14E+06	1.17E+06	1.13E+06	1.17E+06
Citrate_M1_F0	1.36E+04	1.32E+04	1.65E+04	1.43E+04	1.99E+04	1.62E+04	1.78E+04	2.22E+04	1.44E+04	1.41E+04	1.42E+04	1.47E+04
Citrate_M1_F1	7.65E+04	8.29E+04	7.14E+04	8.48E+04	7.80E+04	7.72E+04	7.99E+04	8.94E+04	8.34E+04	7.96E+04	7.21E+04	9.32E+04
Citrate_M2_F1	1.58E+05	1.47E+05	1.37E+05	1.32E+05	1.51E+05	1.32E+05	3.85E+04	2.98E+04	3.48E+04	3.27E+04	3.42E+04	3.86E+04
Citrate_M2_F2	2.34E+05	2.39E+05	2.41E+05	2.78E+05	2.65E+05	2.22E+05	6.41E+04	6.32E+04	7.46E+04	6.55E+04	7.01E+04	7.93E+04
Citrate_M3_F2	9.91E+04	9.16E+04	9.65E+04	1.10E+05	1.18E+05	1.00E+05	1.62E+04	1.33E+04	1.49E+04	1.68E+04	1.68E+04	1.79E+04
Citrate_M3_F3	2.73E+05	2.41E+05	2.81E+05	3.02E+05	2.95E+05	2.40E+05	4.37E+04	4.82E+04	4.78E+04	4.67E+04	4.46E+04	5.01E+04
Citrate_M4_F3	1.09E+05	9.55E+04	9.29E+04	1.14E+05	9.55E+04	8.21E+04	1.14E+04	8.25E+03	1.23E+04	1.44E+04	1.10E+04	1.47E+04
Citrate_M4_F4	6.74E+04	6.97E+04	7.16E+04	8.07E+04	8.79E+04	8.36E+04	1.15E+04	3.05E+03	1.20E+04	8.16E+03	8.63E+03	1.74E+04
Citrate_M5_F4	3.07E+05	2.58E+05	2.84E+05	3.53E+05	3.25E+05	2.83E+05	1.68E+04	1.05E+04	1.97E+04	1.47E+04	1.97E+04	2.68E+04
Citrate_M5_F5	1.09E+05	1.03E+05	9.90E+04	1.25E+05	1.20E+05	1.12E+05	5.78E+03	4.10E+03	9.39E+03	3.99E+03	9.18E+03	1.17E+04
Citrate_M6_F5	3.59E+04	3.09E+04	2.72E+04	4.12E+04	3.11E+04	3.30E+04	1.97E+03	N/A	3.56E+03	N/A	2.44E+03	5.85E+03
Succinate_M0	5.23E+05	1.06E+06	5.03E+05	4.16E+05	6.00E+05	5.92E+05	7.80E+05	7.37E+05	5.50E+05	6.11E+05	5.92E+05	6.40E+05
Succinate_M1_F0	9.96E+03	1.27E+04	8.63E+03	6.98E+03	7.64E+03	8.41E+03	1.14E+04	1.20E+04	8.51E+03	8.93E+03	1.03E+04	1.20E+04
Succinate_M1_F1	2.07E+04	3.86E+04	2.10E+04	1.74E+04	2.35E+04	2.05E+04	3.09E+04	3.73E+04	2.69E+04	2.53E+04	3.03E+04	3.31E+04
Succinate_M2_F1	2.63E+04	3.02E+04	2.98E+04	2.71E+04	3.13E+04	2.77E+04	1.39E+04	1.79E+04	1.52E+04	1.57E+04	2.31E+04	2.56E+04
Succinate_M2_F2	2.72E+04	3.81E+04	2.44E+04	2.91E+04	3.31E+04	3.21E+04	2.80E+04	1.75E+04	2.15E+04	1.88E+04	2.35E+04	3.31E+04
Succinate_M3_F2	5.53E+03	6.37E+03	6.60E+03	7.39E+03	6.68E+03	7.03E+03	2.19E+03	3.20E+03	2.92E+03	1.94E+03	3.80E+03	6.18E+03
Succinate_M3_F3	3.61E+03	4.14E+03	3.84E+03	4.29E+03	4.73E+03	3.68E+03	1.38E+03	N/A	1.83E+03	1.65E+03	2.38E+03	3.67E+03
Succinate_M4_F3	4.08E+04	4.79E+04	4.15E+04	4.60E+04	4.73E+04	4.45E+04	5.74E+03	7.11E+03	9.67E+03	7.56E+03	1.27E+04	2.28E+04
Lactate_M0	2.26E+06	2.18E+06	2.04E+06	1.41E+06	2.11E+06	2.76E+06	2.37E+06	1.05E+06	1.63E+06	2.65E+06	2.94E+06	1.68E+06
Lactate_M1_F0	2.93E+04	3.00E+04	3.00E+04	2.02E+04	3.04E+04	3.73E+04	3.17E+04	1.44E+04	2.11E+04	3.39E+04	3.82E+04	2.16E+04
Lactate_M1_F1	5.91E+04	5.92E+04	5.56E+04	4.13E+04	5.74E+04	7.34E+04	6.02E+04	2.83E+04	4.24E+04	6.85E+04	7.70E+04	4.42E+04
Lactate_M2_F1	3.48E+04	4.83E+04	4.35E+04	4.89E+04	5.07E+04	4.55E+04	6.03E+03	5.98E+03	7.51E+03	6.69E+03	7.70E+03	9.03E+03
Lactate_M2_F2	1.06E+05	1.47E+05	1.27E+05	1.51E+05	1.47E+05	1.33E+05	1.80E+04	1.84E+04	2.56E+04	2.33E+04	2.79E+04	3.10E+04
Lactate_M3_F2	1.32E+06	1.85E+06	1.62E+06	1.85E+06	1.90E+06	1.72E+06	1.19E+05	1.34E+05	1.77E+05	1.51E+05	1.92E+05	2.48E+05
Aspartate_M0	4.09E+06	5.21E+06	5.60E+06	4.60E+06	5.26E+06	4.58E+06	8.02E+06	9.01E+06	1.17E+07	7.56E+06	7.31E+06	7.27E+06
Aspartate_M1_F0	6.34E+04	8.62E+04	8.66E+04	7.59E+04	8.16E+04	7.14E+04	1.22E+05	1.49E+05	1.74E+05	1.14E+05	1.16E+05	1.22E+05
Aspartate_M1_F1	2.02E+05	2.75E+05	2.93E+05	2.53E+05	2.72E+05	2.31E+05	4.15E+05	5.01E+05	5.74E+05	3.99E+05	3.92E+05	3.89E+05
Aspartate_M2_F1	1.28E+05	1.82E+05	1.76E+05	1.78E+05	1.69E+05	1.55E+05	1.49E+05	1.50E+05	1.65E+05	1.44E+05	1.60E+05	1.59E+05
Aspartate_M2_F2	1.15E+06	1.55E+06	1.45E+06	1.50E+06	1.40E+06	1.39E+06	4.27E+05	4.32E+05	4.93E+05	3.76E+05	4.38E+05	4.51E+05
Aspartate_M3_F2	2.32E+06	3.01E+06	2.93E+06	2.90E+06	2.84E+06	2.76E+06	4.85E+05	5.05E+05	6.09E+05	4.68E+05	5.41E+05	5.30E+05
Aspartate_M3_F3	2.30E+06	3.18E+06	3.09E+06	3.04E+06	2.87E+06	2.78E+06	5.43E+05	5.14E+05	6.01E+05	4.87E+05	5.57E+05	5.18E+05
Aspartate_M4_F3	2.07E+05	2.50E+05	2.46E+05	2.68E+05	2.37E+05	2.34E+05	4.74E+04	4.32E+04	5.84E+04	4.53E+04	5.56E+04	5.40E+04
Glutamate_M0	1.50E+07	1.88E+07	1.74E+07	1.84E+07	1.84E+07	1.79E+07	2.86E+07	2.89E+07	3.10E+07	2.69E+07	2.96E+07	2.96E+07
Glutamate_M1_F0	1.98E+05	2.50E+05	2.48E+05	2.32E+05	2.31E+05	2.42E+05	3.78E+05	3.70E+05	4.07E+05	3.59E+05	3.79E+05	4.01E+05
Glutamate_M1_F1	8.93E+05	1.13E+06	1.09E+06	1.06E+06	1.08E+06	1.07E+06	1.90E+06	1.89E+06	1.98E+06	1.85E+06	2.00E+06	2.00E+06
Glutamate_M2_F1	1.36E+06	1.64E+06	1.52E+06	1.68E+06	1.42E+06	1.62E+06	6.27E+05	5.79E+05	7.12E+05	5.85E+05	8.16E+05	7.45E+05
Glutamate_M2_F2	1.85E+06	2.28E+06	2.18E+06	2.35E+06	2.13E+06	2.35E+06	1.74E+06	1.52E+06	1.86E+06	1.63E+06	2.16E+06	2.00E+06
Glutamate_M3_F2	2.58E+05	2.98E+05	2.89E+05	3.14E+05	2.80E+05	3.16E+05	1.36E+05	1.20E+05	1.51E+05	1.33E+05	1.77E+05	1.70E+05
Glutamate_M3_F3	1.06E+06	1.23E+06	1.22E+06	1.36E+06	1.24E+06	1.36E+06	6.39E+05	5.45E+05	6.79E+05	6.04E+05	7.92E+05	7.78E+05
Glutamate_M4_F3	1.72E+06	1.94E+06	1.75E+06	2.32E+06	1.78E+06	2.16E+06	2.13E+05	1.78E+05	3.25E+05	2.13E+05	3.45E+05	3.54E+05
Glutamate_M4_F4	1.83E+05	1.95E+05	1.81E+05	2.38E+05	1.86E+05	2.14E+05	3.52E+04	2.93E+04	4.53E+04	3.40E+04	5.11E+04	4.77E+04
Glutamate_M5_F4	1.02E+06	1.11E+06	1.09E+06	1.38E+06	1.08E+06	1.26E+06	1.05E+05	8.76E+04	1.55E+05	1.04E+05	1.59E+05	1.68E+05
Phosphoenolpyruvate_M0	1.94E+04	1.82E+04	2.01E+04	1.05E+04	1.08E+04	1.23E+04	5.75E+04	6.89E+04	6.33E+04	5.40E+04	5.24E+04	6.15E+04
Phosphoenolpyruvate_M1_F0	3.06E+03	1.38E+03	2.08E+03	1.48E+03	N/A	2.46E+03	2.62E+03	3.92E+03	5.09E+03	1.94E+03	4.12E+03	1.83E+03
Phosphoenolpyruvate_M2_F0	1.21E+04	1.21E+04	1.35E+04	7.72E+03	9.76E+03	7.27E+03	6.78E+03	2.87E+03	2.49E+03	4.97E+03	N/A	5.98E+03
Phosphoenolpyruvate_M3_F0	2.43E+05	2.17E+05	2.87E+05	1.63E+05	1.86E+05	1.90E+05	5.02E+04	4.73E+04	6.07E+04	5.53E+04	6.08E+04	6.62E+04
Glucose6-Phosphate_M0	3.27E+04	5.07E+04	4.83E+04	3.82E+04	3.44E+04	4.76E+04	3.32E+04	3.79E+04	3.61E+04	3.03E+04	2.85E+04	3.06E+04
Glucose6-Phosphate_M5	5.97E+04	2.03E+04	7.84E+04	1.60E+04	2.11E+04	2.38E+04	8.59E+03	6.72E+03	1.08E+04	7.96E+03	9.80E+03	1.05E+04
Glucose6-Phosphate_M6	8.71E+05	2.59E+05	1.04E+06	2.78E+05	2.93E+05	2.85E+05	9.78E+04	7.80E+04	1.26E+05	1.03E+05	1.22E+05	1.48E+05
Malate_M0	4.91E+06	8.73E+06	5.97E+06	5.59E+06	6.46E+06	5.46E+06	1.85E+06	2.27E+06	2.53E+06	1.85E+06	1.78E+06	1.92E+06
Malate_M1_F1	3.36E+05	5.02E+05	3.98E+05	3.85E+05	4.02E+05	3.48E+05	1.21E+05	1.46E+05	1.64E+05	1.28E+05	1.19E+05	1.45E+05
Malate_M2_F2	1.72E+06	2.09E+06	1.92E+06	2.28E+06	2.17E+06	1.99E+06	1.80E+05	1.75E+05	2.26E+05	1.67E+05	1.99E+05	2.23E+05
Malate_M3_F3	5.25E+06	7.03E+06	6.35E+06	7.83E+06	7.68E+06	6.58E+06	3.47E+05	3.86E+05	4.85E+05	4.02E+05	4.30E+05	4.60E+05
Malate_M4_F4	5.42E+05	5.35E+05	5.90E+05	6.16E+05	5.65E+05	5.39E+05	3.39E+04	3.93E+04	6.19E+04	4.16E+04	3.58E+04	5.57E+04
Glutamine_M0	1.53E+07	1.60E+07	1.61E+07	1.76E+07	1.84E+07	1.78E+07	1.74E+07	1.82E+07	4.89E+07	2.08E+07	1.94E+07	2.15E+07
Glutamine_M1_F1	9.45E+05	1.06E+06	1.08E+06	1.14E+06	1.25E+06	1.12E+06	1.12E+06	1.15E+06	3.46E+06	1.41E+06	1.22E+06	1.33E+06
Glutamine_M2_F2	1.39E+05	1.59E+05	1.58E+05	1.80E+05	1.90E+05	1.52E+05	1.25E+05	1.38E+05	3.26E+05	1.55E+05	1.63E+05	1.59E+05
Glutamine_M3_F3	3.49E+04	3.73E+04	4.25E+04	4.66E+04	2.93E+04	3.44E+04	1.63E+04	3.02E+04	2.62E+04	5.42E+04	3.34E+04	2.82E+04
Glutamine_M4_F4	4.85E+04	5.10E+04	4.67E+04	6.43E+04	5.43E+04	4.70E+04	N/A	N/A	N/A	N/A	N/A	N/A
Glutamine_M5_F5	2.48E+04	2.41E+04	2.55E+04	2.75E+04	2.06E+04	2.31E+04	2.71E+03	N/A	N/A	N/A	N/A	3.20E+03
Glycerol3-Phosphate_M0	2.36E+06	2.76E+06	2.68E+06	3.10E+06	2.79E+06	2.53E+06	6.95E+05	8.43E+05	7.77E+05	6.45E+05	6.87E+05	6.60E+05
Glycerol3-Phosphate_M1	2.08E+05	2.10E+05	2.49E+05	2.74E+05	2.30E+05	2.36E+05	5.13E+04	5.52E+04	5.34E+04	4.84E+04	4.60E+04	4.40E+04
Glycerol3-Phosphate_M2	1.33E+06	1.56E+06	1.53E+06	1.83E+06	1.67E+06	1.71E+06	6.79E+04	6.39E+04	9.06E+04	6.98E+04	9.75E+04	7.81E+04
Glycerol3-Phosphate_M3	2.65E+07	2.95E+07	2.73E+07	3.50E+07	3.33E+07	3.26E+07	7.61E+05	7.38E+05	9.76E+05	7.20E+05	1.07E+06	1.12E+06
AcetylCoA_M0	3.19E+05	3.95E+05	3.91E+05	3.99E+05	3.62E+05	3.61E+05	4.65E+05	5.54E+05	4.51E+05	4.45E+05	3.77E+05	3.58E+05
AcetylCoA_M1	1.07E+05	1.33E+05	1.32E+05	1.30E+05	1.21E+05	1.19E+05	1.50E+05	1.81E+05	1.43E+05	1.45E+05	1.28E+05	1.16E+05
AcetylCoA_M2	2.27E+05	2.48E+05	2.56E+05	2.88E+05	2.27E+05	2.53E+05	8.54E+04	9.61E+04	8.07E+04	7.98E+04	7.33E+04	6.79E+04
Cis-Aconitate_M0	38190	50690	39330	35810	44080	42420	42940	48490	40770	45680	49200	50270
Cis-Aconitate_M2_F1	15750	17730	15990	17740	16560	14480	5361	3318	5994	4568	4634	5441
Cis-Aconitate_M3_F2	15220	10090	11900	12280	12080	10720	N/A	2971	5437	3050	1726	2564
Cis-Aconitate_M3_F3	11530	9721	14310	12210	13870	14200	N/A	6633	2702	3486	3944	3509
Cis-Aconitate_M4_F3	7959	10030	7271	9962	10110	8557	641.2	556.9	918.5	1025	1097	1682
Cis-Aconitate_M5_F4	12980	11980	10700	18080	13580	13390	N/A	866.5	N/A	N/A	N/A	1711
Cis-Aconitate_M6_F4	2238	2847	2544	2333	1992	2837	N/A	N/A	N/A	N/A	N/A	748
MS_METABOLITE_DATA_END
#METABOLITES
METABOLITES_START
metabolite_name	m/z	PubChem ID	KEGG_ID
Citrate_M0	191	311	C00158
Citrate_M1_F0	192	311	C00158
Citrate_M1_F1	192	311	C00158
Citrate_M2_F1	193	311	C00158
Citrate_M2_F2	193	311	C00158
Citrate_M3_F2	194	311	C00158
Citrate_M3_F3	194	311	C00158
Citrate_M4_F3	195	311	C00158
Citrate_M4_F4	195	311	C00158
Citrate_M5_F4	196	311	C00158
Citrate_M5_F5	196	311	C00158
Citrate_M6_F5	197	311	C00158
Succinate_M0	117	1110	C00042
Succinate_M1_F0	118	1110	C00042
Succinate_M1_F1	118	1110	C00042
Succinate_M2_F1	119	1110	C00042
Succinate_M2_F2	119	1110	C00042
Succinate_M3_F2	120	1110	C00042
Succinate_M3_F3	120	1110	C00042
Succinate_M4_F3	121	1110	C00042
Lactate_M0	89	91435	C00186
Lactate_M1_F0	90	91435	C00186
Lactate_M1_F1	90	91435	C00186
Lactate_M2_F1	91	91435	C00186
Lactate_M2_F2	91	91435	C00186
Lactate_M3_F2	92	91435	C00186
Aspartate_M0	132	5960	C00049
Aspartate_M1_F0	133	5960	C00049
Aspartate_M1_F1	133	5960	C00049
Aspartate_M2_F1	134	5960	C00049
Aspartate_M2_F2	134	5960	C00049
Aspartate_M3_F2	135	5960	C00049
Aspartate_M3_F3	135	5960	C00049
Aspartate_M4_F3	136	5960	C00049
Glutamate_M0	146	33032	C00025
Glutamate_M1_F0	147	33032	C00025
Glutamate_M1_F1	147	33032	C00025
Glutamate_M2_F1	148	33032	C00025
Glutamate_M2_F2	148	33032	C00025
Glutamate_M3_F2	149	33032	C00025
Glutamate_M3_F3	149	33032	C00025
Glutamate_M4_F3	150	33032	C00025
Glutamate_M4_F4	150	33032	C00025
Glutamate_M5_F4	151	33032	C00025
Phosphoenolpyruvate_M0	167	1005	C00074
Phosphoenolpyruvate_M1_F0	168	1005	C00074
Phosphoenolpyruvate_M2_F0	169	1005	C00074
Phosphoenolpyruvate_M3_F0	170	1005	C00074
Glucose6-Phosphate_M0	259	5958	C00668 
Glucose6-Phosphate_M5	264	5958	C00668 
Glucose6-Phosphate_M6	265	5958	C00668 
Malate_M0	133	525	C00711
Malate_M1_F1	134	525	C00711
Malate_M2_F2	135	525	C00711
Malate_M3_F3	136	525	C00711
Malate_M4_F4	137	525	C00711
Glutamine_M0	145	5961	C00064
Glutamine_M1_F1	146	5961	C00064
Glutamine_M2_F2	147	5961	C00064
Glutamine_M3_F3	148	5961	C00064
Glutamine_M4_F4	149	5961	C00064
Glutamine_M5_F5	150	5961	C00064
Glycerol3-Phosphate_M0	171	439162	C00093
Glycerol3-Phosphate_M1	172	439162	C00093
Glycerol3-Phosphate_M2	173	439162	C00093
Glycerol3-Phosphate_M3	174	439162	C00093
AcetylCoA_M0	808	444493	C00024
AcetylCoA_M1	809	444493	C00024
AcetylCoA_M2	810	444493	C00024
Cis-Aconitate_M0	173	643757	C00417 
Cis-Aconitate_M2_F1	175	643757	C00417 
Cis-Aconitate_M3_F2	176	643757	C00417 
Cis-Aconitate_M3_F3	176	643757	C00417 
Cis-Aconitate_M4_F3	177	643757	C00417 
Cis-Aconitate_M5_F4	178	643757	C00417 
Cis-Aconitate_M6_F4	179	643757	C00417 
METABOLITES_END
#END