Summary of Study ST002930

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 PR001822. The data can be accessed directly via it's Project DOI: 10.21228/M86H8Z This work is supported by NIH grant, U2C- DK119886.

See: https://www.metabolomicsworkbench.org/about/howtocite.php

This study contains a large results data set and is not available in the mwTab file. It is only available for download via FTP as data file(s) here.

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Study IDST002930
Study TitleRole of Hypoxia-inducible factor-1a (HIF1a)in Skeletal Muscle Physiology (C2C12 myotube model)
Study SummaryHypoxia-inducible factor (HIF)-1a is continuously synthesized and degraded in normoxia, whereas during hypoxia, HIF1a stabilization restricts cellular oxygen utilization. Less is known about HIF1a function(s) and sex-specific effects during normoxia in the basal state. Since skeletal muscle is the largest protein store in mammals and protein homeostasis has high energy demands, we determined HIF1a function at baseline during normoxia in C2C12 murine myotubes with the use of untargeted metabolomics. 114 samples of extracted metabolites from cells were analyzed using LCMS. We identified that metabolites, especially those in the glycolytic pathway and TCA cycle, were differentially expressed in cells with HIF1a KO compared to WT.
Institute
Cleveland Clinic
DepartmentInflamation and Immunity
LaboratoryDasarathy Lab
Last NameDasarathy
First NameSrinivasan
Address9500 Euclid Avenue
Emaildasaras@ccf.org
Phone2163799846
Submit Date2023-10-05
Raw Data AvailableYes
Raw Data File Type(s)mzXML
Analysis Type DetailLC-MS
Release Date2024-10-28
Release Version1
Srinivasan Dasarathy Srinivasan Dasarathy
https://dx.doi.org/10.21228/M86H8Z
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Project:

Project ID:PR001822
Project DOI:doi: 10.21228/M86H8Z
Project Title:Role of hypoxia inducible factor-1a (HIF1a) in Skeletal Muscle Physiology
Project Summary:Hypoxia-inducible factor (HIF)-1a is continuously synthesized and degraded in normoxia, whereas during hypoxia, HIF1a stabilization restricts cellular oxygen utilization. Less is known about HIF1a function(s) and sex-specific effects during normoxia in the basal state. Since skeletal muscle is the largest protein store in mammals and protein homeostasis has high energy demands, we determined HIF1a function at baseline during normoxia in skeletal muscle with the use of untargeted metabolomics.
Institute:Cleveland Clinic
Department:Inflammation and Immunity
Last Name:Dasarathy
First Name:Srinivasan
Address:9500 Euclid Avenue
Email:dasaras@ccf.org
Phone:2163799846
Funding Source:NIH

Subject:

Subject ID:SU003043
Subject Type:Cultured cells
Subject Species:Mus musculus
Taxonomy ID:10090
Age Or Age Range:Passage 4-8
Gender:Female

Factors:

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

mb_sample_id local_sample_id Genotype Treatment
SA318096HIF KO C2C12 UnT3 M_Dasarathy_Nicole_114Cells_02042022_HILIC_NEG251HIF1a KO Untreated
SA318097HIF KO C2C12 UnT2 M_Dasarathy_Nicole_114Cells_02042022_HILIC_NEG250HIF1a KO Untreated
SA318098HIF KO C2C12 UnT4 M_Dasarathy_Nicole_114Cells_02042022_HILIC_NEG252HIF1a KO Untreated
SA318099HIF KO C2C12 UnT5 M_Dasarathy_Nicole_114Cells_02042022_HILIC_NEG253HIF1a KO Untreated
SA318100HIF KO C2C12 UnT6 M_Dasarathy_Nicole_114Cells_02042022_HILIC_NEG254HIF1a KO Untreated
SA318101HIF KO C2C12 UnT1 M_Dasarathy_Nicole_114Cells_02042022_HILIC_NEG249HIF1a KO Untreated
SA318102HIF KO C2C12 UnT1 M_Dasarathy_Nicole_114Cells_02042022_HILIC_POS249HIF1a KO Untreated
SA318103HIF KO C2C12 UnT5 M_Dasarathy_Nicole_114Cells_02042022_HILIC_POS253HIF1a KO Untreated
SA318104HIF KO C2C12 UnT6 M_Dasarathy_Nicole_114Cells_02042022_HILIC_POS254HIF1a KO Untreated
SA318105HIF KO C2C12 UnT4 M_Dasarathy_Nicole_114Cells_02042022_HILIC_POS252HIF1a KO Untreated
SA318106HIF KO C2C12 UnT3 M_Dasarathy_Nicole_114Cells_02042022_HILIC_POS251HIF1a KO Untreated
SA318107HIF KO C2C12 UnT2 M_Dasarathy_Nicole_114Cells_02042022_HILIC_POS250HIF1a KO Untreated
SA318108WT C2C12 UnT7 M_Dasarathy_Nicole_114Cells_02042022_HILIC_POS204Wild-type Untreated
SA318109WT C2C12 UnT6 M_Dasarathy_Nicole_114Cells_02042022_HILIC_POS203Wild-type Untreated
SA318110WT C2C12 UnT9 M_Dasarathy_Nicole_114Cells_02042022_HILIC_POS206Wild-type Untreated
SA318111WT C2C12 UnT5 M_Dasarathy_Nicole_114Cells_02042022_HILIC_POS202Wild-type Untreated
SA318112WT C2C12 UnT8 M_Dasarathy_Nicole_114Cells_02042022_HILIC_POS205Wild-type Untreated
SA318113WT C2C12 unit1 M_Dasarathy_Nicole_114Cells_02042022_HILIC_POS189Wild-type Untreated
SA318114WT C2C12 UnT5 M_Dasarathy_Nicole_114Cells_02042022_HILIC_NEG202Wild-type Untreated
SA318115WT C2C12 UnT6 M_Dasarathy_Nicole_114Cells_02042022_HILIC_NEG203Wild-type Untreated
SA318116WT C2C12 UnT4 M_Dasarathy_Nicole_114Cells_02042022_HILIC_NEG201Wild-type Untreated
SA318117WT C2C12 unit3 M_Dasarathy_Nicole_114Cells_02042022_HILIC_NEG191Wild-type Untreated
SA318118WT C2C12 unit2 M_Dasarathy_Nicole_114Cells_02042022_HILIC_NEG190Wild-type Untreated
SA318119WT C2C12 UnT7 M_Dasarathy_Nicole_114Cells_02042022_HILIC_NEG204Wild-type Untreated
SA318120WT C2C12 UnT8 M_Dasarathy_Nicole_114Cells_02042022_HILIC_NEG205Wild-type Untreated
SA318121WT C2C12 unit3 M_Dasarathy_Nicole_114Cells_02042022_HILIC_POS191Wild-type Untreated
SA318122WT C2C12 unit2 M_Dasarathy_Nicole_114Cells_02042022_HILIC_POS190Wild-type Untreated
SA318123WT C2C12 unit1 M_Dasarathy_Nicole_114Cells_02042022_HILIC_NEG189Wild-type Untreated
SA318124WT C2C12 UnT9 M_Dasarathy_Nicole_114Cells_02042022_HILIC_NEG206Wild-type Untreated
SA318125WT C2C12 UnT4 M_Dasarathy_Nicole_114Cells_02042022_HILIC_POS201Wild-type Untreated
Showing results 1 to 30 of 30

Collection:

Collection ID:CO003036
Collection Summary:Metabolites were extracted from myotubes and skeletal muscle using chilled methanol containing 6 labeled internal standards (Betaine-d9, Carnitine-d9, Estrone-13C3, Cholesterol-13C3, Valeric acid-d9, Choline-d13). After centrifugation at 14,000g for 20 minutes to precipitate the protein pellet, the supernatant was removed, dried, and reconstituted in 5% acetonitrile. LC/MS (liquid chromatography/mass spectrometry) analysis was then performed, including on pooled quality control samples.
Sample Type:Skeletal myotubes

Treatment:

Treatment ID:TR003052
Treatment Summary:Cells were not treated (Untreated, UnT)

Sample Preparation:

Sampleprep ID:SP003049
Sampleprep Summary:Metabolites were extracted from myotubes and skeletal muscle using chilled methanol containing 6 labeled internal standards (Betaine-d9, Carnitine-d9, Estrone-13C3, Cholesterol-13C3, Valeric acid-d9, Choline-d13). After centrifugation at 14,000g for 20 minutes to precipitate the protein pellet, the supernatant was removed, dried, and reconstituted in 5% acetonitrile. LC/MS (liquid chromatography/mass spectrometry) analysis was then performed, including on pooled quality control samples.

Combined analysis:

Analysis ID AN004807 AN004808
Analysis type MS MS
Chromatography type HILIC HILIC
Chromatography system Thermo Vanquish Thermo Vanquish
Column Waters ACQUITY UPLC BEH Amide (150 x 2.1mm,1.7um) Waters ACQUITY UPLC BEH Amide (150 x 2.1mm,1.7um)
MS Type ESI ESI
MS instrument type QTRAP QTRAP
MS instrument name Thermo Q Exactive Focus Thermo Q Exactive Focus
Ion Mode POSITIVE NEGATIVE
Units relative abundance relative abundance

Chromatography:

Chromatography ID:CH003633
Instrument Name:Thermo Vanquish
Column Name:Waters ACQUITY UPLC BEH Amide (150 x 2.1mm,1.7um)
Column Temperature:40
Flow Gradient:Time (min.) Flow Rate (mL/min) %B 0.0 0.2 100 2.0 0.2 70 4.0 0.2 60 5.5 0.2 60 10.5 0.2 50 11.5 0.2 50 12.0 0.2 100 16.0 0.2 100
Flow Rate:0.2 mL/min
Solvent A:100% water;10 mM ammonium acetate; 0.125% acetic acid
Solvent B:5% water/95% acetonitrile;10 mM ammonium acetate; 0.125% acetic acid
Chromatography Type:HILIC

MS:

MS ID:MS004553
Analysis ID:AN004807
Instrument Name:Thermo Q Exactive Focus
Instrument Type:QTRAP
MS Type:ESI
MS Comments:Data dependent acquisitions (DDA) on the pooled representative QC samples include MS full scans at a resolution of 120,000 and HCD MS/MS scans taken on the top 5 most abundant ions at a resolution of 30,000 with dynamic exclusion. The resolution of the MS2 scans were taken at a stepped NCE energy of 10.0, 20.0 and 30.0.
Ion Mode:POSITIVE
  
MS ID:MS004554
Analysis ID:AN004808
Instrument Name:Thermo Q Exactive Focus
Instrument Type:QTRAP
MS Type:ESI
MS Comments:Data dependent acquisitions (DDA) on the pooled representative QC samples include MS full scans at a resolution of 120,000 and HCD MS/MS scans taken on the top 5 most abundant ions at a resolution of 30,000 with dynamic exclusion. The resolution of the MS2 scans were taken at a stepped NCE energy of 10.0, 20.0 and 30.0.
Ion Mode:NEGATIVE
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