Summary of Study ST001178

This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR000790. The data can be accessed directly via it's Project DOI: 10.21228/M8K40J This work is supported by NIH grant, U2C- DK119886.

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Study ID	ST001178
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
Submit Date	2019-04-22
Analysis Type Detail	CE-MS
Release Date	2020-03-03
Release Version	1

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Sample Preparation:

Sampleprep ID:	SP001252
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.

Sampleprep ID:

SP001252

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.