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Series GSE6766 Query DataSets for GSE6766
Status Public on Jan 18, 2007
Title C2C12 Myotubes in response to Palmitate
Organism Mus musculus
Experiment type Expression profiling by array
Summary To identify mediators of obesity-linked reductions in PGC-1, we tested the effects of cellular nutrients in C2C12 myotubes. While overnight exposure to high insulin, glucose, glucosamine, or amino acids had no effect, saturated fatty acids (FA) potently reduced PGC-1a and b mRNA expression.
Keywords: Nutrient Effect
 
Overall design Cell culture - Mouse C2C12 myoblasts (ATCC, Manassas, VA) were maintained in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 20% fetal bovine serum (Invitrogen), at a confluency of 60-70%. To initiate differentiation, cells were allowed to reach 100% confluency and medium was changed to DMEM containing 2% horse serum (Invitrogen) and changed every 2 days. Full differentiation, with myotube fusion and spontaneous twitching, was observed at 5 days. Fatty acid stock preparation - Fatty acids were dissolved in 0.1 N sodium hydroxide (final concentration 100 mM) at 65°C for 2 hours and then complexed with 10% fatty acid-free BSA, yielding a final stock of 5 mM. Three replicates for each fatty-acid. Microarray analysis - RNA was isolated as described from C2C12 myotubes treated overnight with 500 µM palmitate or 1% BSA, and cRNA was synthesized. 10 mg of cRNA were hybridized to Affymetrix mouse 430A 2.0 arrays. Intensity values were quantified using MAS 5.0 software. MAPPFinder (www.genmapp.org) was used to integrate expression data with known pathways.
 
Contributor(s) Crunkhorn S, Patti M
Citation(s) 17416903
Submission date Jan 17, 2007
Last update date May 04, 2018
Contact name Sarah Crunkhorn
E-mail(s) Sarah.Crunkhorn@joslin.harvard.edu
Phone 61726424795
URL http://www.joslinresearch.org
Organization name Joslin Diabetes Center
Department CMP
Lab Patti Lab
Street address 1 Joslin Place
City boston
State/province MA
ZIP/Postal code 02215
Country USA
 
Platforms (1)
GPL8321 [Mouse430A_2] Affymetrix Mouse Genome 430A 2.0 Array
Samples (6)
GSM152022 C2C12 Myotubes treated with BSA, rep1
GSM152023 C2C12 Myotubes treated with BSA, rep2
GSM152024 C2C12 Myotubes treated with BSA, rep3
Relations
BioProject PRJNA98939

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Supplementary file Size Download File type/resource
GSE6766_RAW.tar 11.6 Mb (http)(custom) TAR (of CEL)
Raw data provided as supplementary file

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