{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST001178","ANALYSIS_ID":"AN001955","VERSION":"1","CREATED_ON":"May 1, 2019, 2:29 pm"},

"PROJECT":{"PROJECT_TITLE":"Metabolomic analysis of C2C12 myoblasts induced by the transcriptional factor FOXO1","PROJECT_SUMMARY":"To determine the metabolic changes occurring due to the activation of transcriptional factor FOXO1, we analyzed the metabolic profile of C2C12 myoblasts expressing FOXO1-estrogen receptor fusion protein using CE-TOFMS.","INSTITUTE":"Kyoto Prefectural University","LAST_NAME":"Kamei","FIRST_NAME":"Yasutomi","ADDRESS":"1-5 Hangi-cho, Shimogamo, Sakyo-ku Kyoto, Kyoto, Kyoto, 606-8522, Japan","EMAIL":"cnqwb974@yahoo.co.jp","PHONE":"+81-75-703-5661"},

"STUDY":{"STUDY_TITLE":"Metabolomic analysis of C2C12 myoblasts induced by the transcriptional factor FOXO1","STUDY_SUMMARY":"The transcriptional factor FOXO1 is considered to play roles in the regulation of energy metabolism in various tissues. To determine the metabolic changes occurring due to the activation of FOXO1, we analyzed the metabolic profile of C2C12 myoblasts expressing FOXO1-estrogen receptor fusion protein using CE-TOFMS. In the FOXO1-activated cells, the metabolite levels during glycolysis were higher. In addition, the gene expression of pyruvate dehydrogenase kinase, an enzyme that inhibits glucose utilization, increased. In the FOXO1-activated cells, the metabolite levels of numerous amino acids decreased, with increased gene expression of branched chain amino acid metabolism enzymes. Our results suggest that FOXO1 suppresses glucose utilization and promotes the use of proteins/amino acids as energy sources in muscle cells, potentially during starvation.","INSTITUTE":"Kyoto Prefectural University","LAST_NAME":"Kamei","FIRST_NAME":"Yasutomi","ADDRESS":"1-5 Hangi-cho, Shimogamo, Sakyo-ku Kyoto, Kyoto, Kyoto, 606-8522, Japan","EMAIL":"cnqwb974@yahoo.co.jp","PHONE":"+81-75-703-5661"},

"SUBJECT":{"SUBJECT_TYPE":"Cultured cells","SUBJECT_SPECIES":"Mus musculus","TAXONOMY_ID":"10090"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"-",
"Sample ID":"Control 1",
"Factors":{"treatment":"control"},
"Additional sample data":{"Hours":"24","tamoxifen(µM）":"-","Cells(×106cells)":"3.9"}
},
{
"Subject ID":"-",
"Sample ID":"Control 2",
"Factors":{"treatment":"control"},
"Additional sample data":{"Hours":"24","tamoxifen(µM）":"-","Cells(×106cells)":"3.9"}
},
{
"Subject ID":"-",
"Sample ID":"Control 3",
"Factors":{"treatment":"control"},
"Additional sample data":{"Hours":"24","tamoxifen(µM）":"-","Cells(×106cells)":"3.9"}
},
{
"Subject ID":"-",
"Sample ID":"FOXO1-ER1",
"Factors":{"treatment":"tamoxifen"},
"Additional sample data":{"Hours":"24","tamoxifen(µM）":"2","Cells(×106cells)":"3.7"}
},
{
"Subject ID":"-",
"Sample ID":"FOXO1-ER2",
"Factors":{"treatment":"tamoxifen"},
"Additional sample data":{"Hours":"24","tamoxifen(µM）":"2","Cells(×106cells)":"3.7"}
},
{
"Subject ID":"-",
"Sample ID":"FOXO1-ER3",
"Factors":{"treatment":"tamoxifen"},
"Additional sample data":{"Hours":"24","tamoxifen(µM）":"2","Cells(×106cells)":"3.7"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"Cells were washed with cold PBS and then flash-frozen in liquid N2.","SAMPLE_TYPE":"Muscle"},

"TREATMENT":{"TREATMENT_SUMMARY":"Two days after confluence, cells were treated with tamoxifen for 24 h and used for the metabolomic analysis."},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"The cells treated with vehicle (control, N = 3) or tamoxifen (FOXO1-ER, N = 3) were used for the metabolomic analysis (Human Metabolome Technologies Inc., Tsuruoka, Japan). 3.9 x 106 cells were transferred into 500 μL of methanol containing 50 mM external standard. After homogenization by BMSM10N21 (BMS, Tokyo) at 1,500 rpm for 120 s performed 5 times, 500 μL of chloroform and 200 μL of ultrapure water were added to the homogenate. The solution was mixed well and centrifuged at 2,300 × g for 5 min at 4 °C. The resultant water phase was ultrafiltered using a Millipore Ultrafree-MC PLHCC HMT Centrifugal Filter Device, 5 kDa (Millipore, Billerica, MA). The filtrate was dried and then dissolved in 50 μL of ultrapure water. The samples obtained were then subjected to capillary electrophoresis time-of-flight mass spectrometry (CE-TOFMS) analysis using the Agilent CE-TOFMS system (Agilent Technologies, Santa Clara, CA) at 4 °C. The detected peaks were aligned according to their m/z values and normalized migration times. The peaks were mean-centered and scaled using their standard deviations on a per peak basis as a pretreatment."},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_TYPE":"None (Direct infusion)","INSTRUMENT_NAME":"none","COLUMN_NAME":"none"},

"ANALYSIS":{"ANALYSIS_TYPE":"MS"},

"MS":{"INSTRUMENT_NAME":"Agilent CE-TOFMS","INSTRUMENT_TYPE":"Other","MS_TYPE":"ESI","ION_MODE":"NEGATIVE","MS_COMMENTS":"-"},

"MS_METABOLITE_DATA":{
"Units":"fold",

"Data":[{"Metabolite":"Pyruvic acid","Control 1":"0.02","Control 2":"0.01","Control 3":"0.01","FOXO1-ER1":"0.01","FOXO1-ER2":"0.01","FOXO1-ER3":"0.01"},{"Metabolite":"Lactic acid","Control 1":"1.30","Control 2":"0.58","Control 3":"0.50","FOXO1-ER1":"1.90","FOXO1-ER2":"1.80","FOXO1-ER3":"1.20"},{"Metabolite":"3-Hydroxybutyric acid","Control 1":"0.00","Control 2":"0.00","Control 3":"","FOXO1-ER1":"0.00","FOXO1-ER2":"0.00","FOXO1-ER3":"0.00"},{"Metabolite":"Glyceric acid","Control 1":"0.00","Control 2":"0.00","Control 3":"","FOXO1-ER1":"0.00","FOXO1-ER2":"0.00","FOXO1-ER3":"0.00"},{"Metabolite":"XA0002","Control 1":"0.02","Control 2":"0.02","Control 3":"0.02","FOXO1-ER1":"0.03","FOXO1-ER2":"0.03","FOXO1-ER3":"0.03"},{"Metabolite":"Fumaric acid","Control 1":"0.01","Control 2":"0.01","Control 3":"0.01","FOXO1-ER1":"0.01","FOXO1-ER2":"0.01","FOXO1-ER3":"0.01"},{"Metabolite":"2-Oxoisovaleric acid","Control 1":"0.01","Control 2":"0.00","Control 3":"0.00","FOXO1-ER1":"0.00","FOXO1-ER2":"0.00","FOXO1-ER3":"0.00"},{"Metabolite":"Hexanoic acid","Control 1":"","Control 2":"","Control 3":"0.00","FOXO1-ER1":"0.00","FOXO1-ER2":"","FOXO1-ER3":"0.00"},{"Metabolite":"Succinic acid","Control 1":"0.01","Control 2":"0.01","Control 3":"0.01","FOXO1-ER1":"0.01","FOXO1-ER2":"0.01","FOXO1-ER3":"0.01"},{"Metabolite":"Isethionic acid","Control 1":"0.00","Control 2":"0.00","Control 3":"0.00","FOXO1-ER1":"0.00","FOXO1-ER2":"0.00","FOXO1-ER3":"0.00"},{"Metabolite":"5-Oxoproline","Control 1":"0.34","Control 2":"0.14","Control 3":"0.13","FOXO1-ER1":"0.30","FOXO1-ER2":"0.30","FOXO1-ER3":"0.23"},{"Metabolite":"4-Methyl-2-oxovaleric acid","Control 1":"0.05","Control 2":"0.02","Control 3":"0.02","FOXO1-ER1":"0.03","FOXO1-ER2":"0.02","FOXO1-ER3":"0.01"},{"Metabolite":"3-Methyl-2-oxovaleric acid","Control 1":"0.05","Control 2":"0.02","Control 3":"0.02","FOXO1-ER1":"0.03","FOXO1-ER2":"0.02","FOXO1-ER3":"0.01"},{"Metabolite":"N-Acetyl-β-alanine","Control 1":"0.00","Control 2":"0.00","Control 3":"0.00","FOXO1-ER1":"0.00","FOXO1-ER2":"0.00","FOXO1-ER3":"0.00"},{"Metabolite":"Malic acid","Control 1":"0.08","Control 2":"0.06","Control 3":"0.07","FOXO1-ER1":"0.11","FOXO1-ER2":"0.13","FOXO1-ER3":"0.11"},{"Metabolite":"Threonic acid","Control 1":"0.03","Control 2":"0.02","Control 3":"0.03","FOXO1-ER1":"0.03","FOXO1-ER2":"0.02","FOXO1-ER3":"0.03"},{"Metabolite":"Ethanolamine phosphate","Control 1":"0.01","Control 2":"0.01","Control 3":"0.01","FOXO1-ER1":"0.01","FOXO1-ER2":"0.01","FOXO1-ER3":"0.01"},{"Metabolite":"Octanoic acid","Control 1":"","Control 2":"","Control 3":"0.00","FOXO1-ER1":"","FOXO1-ER2":"","FOXO1-ER3":"0.00"},{"Metabolite":"XA0004","Control 1":"0.00","Control 2":"0.00","Control 3":"0.00","FOXO1-ER1":"0.00","FOXO1-ER2":"","FOXO1-ER3":"0.00"},{"Metabolite":"2-Oxoglutaric acid","Control 1":"0.03","Control 2":"0.02","Control 3":"0.03","FOXO1-ER1":"0.03","FOXO1-ER2":"0.03","FOXO1-ER3":"0.04"},{"Metabolite":"2-Hydroxyglutaric acid","Control 1":"0.00","Control 2":"0.00","Control 3":"0.00","FOXO1-ER1":"0.00","FOXO1-ER2":"0.00","FOXO1-ER3":"0.00"},{"Metabolite":"Pelargonic acid","Control 1":"0.00","Control 2":"0.00","Control 3":"0.00","FOXO1-ER1":"0.00","FOXO1-ER2":"0.00","FOXO1-ER3":"0.00"},{"Metabolite":"3-Hydroxy-3-methylglutaric acid","Control 1":"0.00","Control 2":"0.00","Control 3":"0.00","FOXO1-ER1":"0.00","FOXO1-ER2":"0.00","FOXO1-ER3":"0.00"},{"Metabolite":"N-Acetylcysteine","Control 1":"0.00","Control 2":"","Control 3":"","FOXO1-ER1":"0.00","FOXO1-ER2":"","FOXO1-ER3":"0.00"},{"Metabolite":"Terephthalic acid","Control 1":"0.00","Control 2":"0.00","Control 3":"0.00","FOXO1-ER1":"0.00","FOXO1-ER2":"0.00","FOXO1-ER3":"0.00"},{"Metabolite":"XA0012","Control 1":"0.00","Control 2":"0.00","Control 3":"0.00","FOXO1-ER1":"0.01","FOXO1-ER2":"0.00","FOXO1-ER3":"0.00"},{"Metabolite":"Phosphoenolpyruvic acid","Control 1":"0.00","Control 2":"0.00","Control 3":"0.00","FOXO1-ER1":"0.00","FOXO1-ER2":"0.01","FOXO1-ER3":"0.01"},{"Metabolite":"Uric acid","Control 1":"0.00","Control 2":"0.00","Control 3":"0.00","FOXO1-ER1":"0.00","FOXO1-ER2":"0.00","FOXO1-ER3":"0.00"},{"Metabolite":"Glyceraldehyde 3-phosphate","Control 1":"0.00","Control 2":"0.00","Control 3":"0.00","FOXO1-ER1":"0.01","FOXO1-ER2":"0.00","FOXO1-ER3":"0.01"},{"Metabolite":"Dihydroxyacetone phosphate","Control 1":"0.01","Control 2":"0.01","Control 3":"0.00","FOXO1-ER1":"0.06","FOXO1-ER2":"0.04","FOXO1-ER3":"0.04"},{"Metabolite":"Glycerol 3-phosphate","Control 1":"0.01","Control 2":"0.01","Control 3":"0.01","FOXO1-ER1":"0.02","FOXO1-ER2":"0.02","FOXO1-ER3":"0.02"},{"Metabolite":"Decanoic acid","Control 1":"0.00","Control 2":"0.00","Control 3":"0.00","FOXO1-ER1":"0.00","FOXO1-ER2":"","FOXO1-ER3":"0.00"},{"Metabolite":"cis-Aconitic acid","Control 1":"0.02","Control 2":"0.02","Control 3":"0.02","FOXO1-ER1":"0.02","FOXO1-ER2":"0.01","FOXO1-ER3":"0.01"},{"Metabolite":"N-Carbamoylaspartic acid","Control 1":"0.00","Control 2":"0.00","Control 3":"0.00","FOXO1-ER1":"","FOXO1-ER2":"","FOXO1-ER3":""},{"Metabolite":"O-Phosphoserine","Control 1":"0.00","Control 2":"0.00","Control 3":"0.00","FOXO1-ER1":"0.00","FOXO1-ER2":"0.00","FOXO1-ER3":"0.00"},{"Metabolite":"3-Phosphoglyceric acid","Control 1":"0.00","Control 2":"0.00","Control 3":"0.00","FOXO1-ER1":"0.01","FOXO1-ER2":"0.01","FOXO1-ER3":"0.01"},{"Metabolite":"2-Phosphoglyceric acid","Control 1":"","Control 2":"0.00","Control 3":"","FOXO1-ER1":"0.00","FOXO1-ER2":"0.00","FOXO1-ER3":"0.00"},{"Metabolite":"N-Acetylglutamine","Control 1":"0.00","Control 2":"0.00","Control 3":"0.00","FOXO1-ER1":"","FOXO1-ER2":"","FOXO1-ER3":""},{"Metabolite":"N-Acetylglutamic acid","Control 1":"0.00","Control 2":"0.00","Control 3":"0.00","FOXO1-ER1":"0.00","FOXO1-ER2":"0.00","FOXO1-ER3":"0.00"},{"Metabolite":"Isocitric acid","Control 1":"0.01","Control 2":"0.01","Control 3":"0.01","FOXO1-ER1":"0.02","FOXO1-ER2":"0.01","FOXO1-ER3":"0.01"},{"Metabolite":"Citric acid","Control 1":"0.27","Control 2":"0.23","Control 3":"0.23","FOXO1-ER1":"0.33","FOXO1-ER2":"0.24","FOXO1-ER3":"0.21"},{"Metabolite":"Quinic acid","Control 1":"","Control 2":"","Control 3":"0.00","FOXO1-ER1":"","FOXO1-ER2":"","FOXO1-ER3":""},{"Metabolite":"Glucuronic acid-1","Control 1":"0.00","Control 2":"0.00","Control 3":"0.00","FOXO1-ER1":"0.00","FOXO1-ER2":"0.00","FOXO1-ER3":"0.00"},{"Metabolite":"Galacturonic acid-1","Control 1":"0.00","Control 2":"0.00","Control 3":"0.00","FOXO1-ER1":"0.00","FOXO1-ER2":"0.00","FOXO1-ER3":"0.00"},{"Metabolite":"Glucuronic acid-2","Control 1":"0.00","Control 2":"0.00","Control 3":"0.00","FOXO1-ER1":"0.00","FOXO1-ER2":"0.00","FOXO1-ER3":"0.00"},{"Metabolite":"Galacturonic acid-2","Control 1":"0.00","Control 2":"0.00","Control 3":"0.00","FOXO1-ER1":"0.00","FOXO1-ER2":"0.00","FOXO1-ER3":"0.00"},{"Metabolite":"Gluconic acid","Control 1":"0.04","Control 2":"0.03","Control 3":"0.04","FOXO1-ER1":"0.09","FOXO1-ER2":"0.06","FOXO1-ER3":"0.07"},{"Metabolite":"Lauric acid","Control 1":"0.02","Control 2":"0.01","Control 3":"0.02","FOXO1-ER1":"0.01","FOXO1-ER2":"0.01","FOXO1-ER3":"0.01"},{"Metabolite":"Phosphocreatine","Control 1":"0.70","Control 2":"0.66","Control 3":"0.72","FOXO1-ER1":"0.55","FOXO1-ER2":"0.59","FOXO1-ER3":"0.56"},{"Metabolite":"O-Succinylhomoserine","Control 1":"0.00","Control 2":"0.00","Control 3":"0.00","FOXO1-ER1":"0.00","FOXO1-ER2":"0.00","FOXO1-ER3":"0.00"},{"Metabolite":"Pantothenic acid","Control 1":"0.04","Control 2":"0.03","Control 3":"0.03","FOXO1-ER1":"0.01","FOXO1-ER2":"0.01","FOXO1-ER3":"0.01"},{"Metabolite":"XA0027","Control 1":"0.00","Control 2":"0.00","Control 3":"0.00","FOXO1-ER1":"0.00","FOXO1-ER2":"0.00","FOXO1-ER3":"0.00"},{"Metabolite":"Ribose 5-phosphate","Control 1":"0.00","Control 2":"0.00","Control 3":"0.00","FOXO1-ER1":"0.01","FOXO1-ER2":"0.01","FOXO1-ER3":"0.01"},{"Metabolite":"Ribulose 5-phosphate","Control 1":"0.01","Control 2":"0.01","Control 3":"0.01","FOXO1-ER1":"0.03","FOXO1-ER2":"0.02","FOXO1-ER3":"0.02"},{"Metabolite":"XA0033","Control 1":"0.17","Control 2":"0.15","Control 3":"0.15","FOXO1-ER1":"0.13","FOXO1-ER2":"0.12","FOXO1-ER3":"0.12"},{"Metabolite":"Glucose 1-phosphate","Control 1":"0.03","Control 2":"0.03","Control 3":"0.03","FOXO1-ER1":"0.06","FOXO1-ER2":"0.05","FOXO1-ER3":"0.05"},{"Metabolite":"Glucose 6-phosphate","Control 1":"0.01","Control 2":"0.01","Control 3":"0.01","FOXO1-ER1":"0.03","FOXO1-ER2":"0.03","FOXO1-ER3":"0.04"},{"Metabolite":"Fructose 6-phosphate","Control 1":"0.00","Control 2":"0.00","Control 3":"0.00","FOXO1-ER1":"0.01","FOXO1-ER2":"0.01","FOXO1-ER3":"0.01"},{"Metabolite":"myo-Inositol 1-phosphate","Control 1":"0.01","Control 2":"0.02","Control 3":"0.02","FOXO1-ER1":"0.02","FOXO1-ER2":"0.03","FOXO1-ER3":"0.02"},{"Metabolite":"myo-Inositol 3-phosphate","Control 1":"0.01","Control 2":"0.02","Control 3":"0.02","FOXO1-ER1":"0.02","FOXO1-ER2":"0.03","FOXO1-ER3":"0.02"},{"Metabolite":"myo-Inositol 2-phosphate","Control 1":"0.00","Control 2":"0.00","Control 3":"0.00","FOXO1-ER1":"0.00","FOXO1-ER2":"0.00","FOXO1-ER3":"0.00"},{"Metabolite":"Sulfotyrosine","Control 1":"0.00","Control 2":"0.00","Control 3":"0.00","FOXO1-ER1":"0.00","FOXO1-ER2":"0.00","FOXO1-ER3":"0.00"},{"Metabolite":"Sorbitol 6-phosphate","Control 1":"0.00","Control 2":"0.00","Control 3":"0.00","FOXO1-ER1":"0.00","FOXO1-ER2":"0.00","FOXO1-ER3":"0.00"},{"Metabolite":"2,3-Diphosphoglyceric acid","Control 1":"0.00","Control 2":"0.00","Control 3":"0.00","FOXO1-ER1":"0.00","FOXO1-ER2":"0.00","FOXO1-ER3":"0.00"},{"Metabolite":"6-Phosphogluconic acid","Control 1":"0.00","Control 2":"0.00","Control 3":"0.00","FOXO1-ER1":"0.00","FOXO1-ER2":"0.00","FOXO1-ER3":"0.00"},{"Metabolite":"Rhein","Control 1":"0.00","Control 2":"0.00","Control 3":"0.00","FOXO1-ER1":"0.00","FOXO1-ER2":"0.00","FOXO1-ER3":"0.00"},{"Metabolite":"Sedoheptulose 7-phosphate","Control 1":"0.01","Control 2":"0.00","Control 3":"0.01","FOXO1-ER1":"0.02","FOXO1-ER2":"0.01","FOXO1-ER3":"0.02"},{"Metabolite":"N-Acetylglucosamine 1-phosphate","Control 1":"0.00","Control 2":"0.00","Control 3":"0.00","FOXO1-ER1":"0.00","FOXO1-ER2":"0.00","FOXO1-ER3":"0.00"},{"Metabolite":"N-Acetylglucosamine 6-phosphate","Control 1":"0.00","Control 2":"0.00","Control 3":"0.00","FOXO1-ER1":"0.00","FOXO1-ER2":"0.00","FOXO1-ER3":"0.00"},{"Metabolite":"N-Acetylneuraminic acid","Control 1":"0.00","Control 2":"0.00","Control 3":"0.00","FOXO1-ER1":"0.00","FOXO1-ER2":"0.00","FOXO1-ER3":"0.00"},{"Metabolite":"Ribulose 1,5-diphosphate","Control 1":"0.00","Control 2":"0.00","Control 3":"0.00","FOXO1-ER1":"0.00","FOXO1-ER2":"0.00","FOXO1-ER3":"0.00"},{"Metabolite":"3'-CMP","Control 1":"0.00","Control 2":"0.00","Control 3":"","FOXO1-ER1":"0.00","FOXO1-ER2":"0.00","FOXO1-ER3":"0.00"},{"Metabolite":"2'-CMP","Control 1":"0.00","Control 2":"0.00","Control 3":"","FOXO1-ER1":"0.00","FOXO1-ER2":"0.00","FOXO1-ER3":"0.00"},{"Metabolite":"CMP","Control 1":"0.00","Control 2":"0.00","Control 3":"0.00","FOXO1-ER1":"0.00","FOXO1-ER2":"0.00","FOXO1-ER3":"0.00"},{"Metabolite":"UMP","Control 1":"0.00","Control 2":"0.00","Control 3":"0.00","FOXO1-ER1":"0.00","FOXO1-ER2":"0.00","FOXO1-ER3":"0.00"},{"Metabolite":"N-Glycolylneuraminic acid","Control 1":"0.00","Control 2":"0.00","Control 3":"0.00","FOXO1-ER1":"0.00","FOXO1-ER2":"0.00","FOXO1-ER3":"0.00"},{"Metabolite":"cAMP","Control 1":"0.00","Control 2":"0.00","Control 3":"0.00","FOXO1-ER1":"","FOXO1-ER2":"","FOXO1-ER3":"0.00"},{"Metabolite":"Phytic acid_divalent","Control 1":"0.01","Control 2":"0.01","Control 3":"0.01","FOXO1-ER1":"0.01","FOXO1-ER2":"0.01","FOXO1-ER3":"0.00"},{"Metabolite":"5-Aminoimidazole-4-carboxamide ribotide","Control 1":"","Control 2":"","Control 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}

}