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 IDST001178
Study TitleMetabolomic analysis of C2C12 myoblasts induced by the transcriptional factor FOXO1
Study SummaryThe 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 NameKamei
First NameYasutomi
Address1-5 Hangi-cho, Shimogamo, Sakyo-ku Kyoto, Kyoto, Kyoto, 606-8522, Japan
Emailcnqwb974@yahoo.co.jp
Phone+81-75-703-5661
Submit Date2019-04-22
Analysis Type DetailLC-MS
Release Date2020-03-03
Release Version1
Yasutomi Kamei Yasutomi Kamei
https://dx.doi.org/10.21228/M8K40J
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Project:

Project ID:PR000790
Project DOI:doi: 10.21228/M8K40J
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

Subject:

Subject ID:SU001244
Subject Type:Cultured cells
Subject Species:Mus musculus
Taxonomy ID:10090

Factors:

Subject type: Cultured cells; Subject species: Mus musculus (Factor headings shown in green)

mb_sample_id local_sample_id treatment
SA081600Control 1control
SA081601Control 3control
SA081602Control 2control
SA081603FOXO1-ER3tamoxifen
SA081604FOXO1-ER1tamoxifen
SA081605FOXO1-ER2tamoxifen
Showing results 1 to 6 of 6

Collection:

Collection ID:CO001238
Collection Summary:Cells were washed with cold PBS and then flash-frozen in liquid N2.
Sample Type:Muscle

Treatment:

Treatment ID:TR001259
Treatment Summary:Two days after confluence, cells were treated with tamoxifen for 24 h and used for the metabolomic analysis.

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.

Combined analysis:

Analysis ID AN001954 AN001955
Analysis type MS MS
Chromatography type None (Direct infusion) None (Direct infusion)
Chromatography system none none
Column none none
MS Type ESI ESI
MS instrument type Other Other
MS instrument name Agilent CE-TOFMS Agilent CE-TOFMS
Ion Mode POSITIVE NEGATIVE
Units Relative Area fold

Chromatography:

Chromatography ID:CH001417
Instrument Name:none
Column Name:none
Chromatography Type:None (Direct infusion)

MS:

MS ID:MS001809
Analysis ID:AN001954
Instrument Name:Agilent CE-TOFMS
Instrument Type:Other
MS Type:ESI
MS Comments:-
Ion Mode:POSITIVE
  
MS ID:MS001810
Analysis ID:AN001955
Instrument Name:Agilent CE-TOFMS
Instrument Type:Other
MS Type:ESI
MS Comments:-
Ion Mode:NEGATIVE
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