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Series GSE30192 Query DataSets for GSE30192
Status Public on Jan 01, 2012
Title Effect of 5-azacytidine on gene expression in C2C12 myoblasts
Organism Mus musculus
Experiment type Expression profiling by array
Summary Mesenchymal progenitor cells can be differentiated in vitro into myotubes that exhibit many characteristic features of primary mammalian skeletal muscle fibers. However, in general, they do not show the functional excitation-contraction coupling or the striated sarcomere arrangement typical of mature myofibers. Epigenetic modifications have been shown to play a key role in regulating the progressional changes in transcription necessary for muscle differentiation. In this study, we demonstrate that treatment of murine C2C12 mesenchymal progenitor cells with 10 µM of the DNA methylation inhibitor 5-azacytidine (5AC) promotes myogenesis, resulting in myotubes with enhanced maturity as compared to untreated myotubes. Specifically, 5AC treatment resulted in the upregulation of muscle genes at the myoblast stage while at later stages nearly 50 % of the 5AC-treated myotubes displayed a mature, well-defined sarcomere organization as well as spontaneous contractions that coincided with action potentials and intracellular calcium transients. Both the percentage of striated myotubes and their contractile activity could be inhibited by 20 nM TTX, 10 µM ryanodine and 100 µM nifedipine, suggesting that action potential-induced calcium transients are responsible for these characteristics. Our data suggest that genomic demethylation induced by 5AC overcomes an epigenetic barrier that prevents untreated C2C12 myotubes from reaching full maturity.
Overall design C2C12 cells were plated at 1500 cells/cm2 (day -1), cultured for 1 day in DMEM 10%NCS, then treated (d0) with or without 10uM 5-azacytidine (5AC) for an additional 3 days. RNA was extracted on day 3 and hybridized to GeneChip Mouse Genome 430 2.0 array (Affymetrix).
Contributor(s) Hupkes M, Sotoca AM, van Someren EP, van Ravestein-van Os RI, Bauerschmidt S, van Zoelen EJ, Dechering KJ
Citation(s) 21795504
Submission date Jun 24, 2011
Last update date Feb 11, 2019
Contact name Marlinda Hupkes
Phone +31(0)243652519
Organization name Radboud University Nijmegen
Department Cell and Applied Biology
Street address Heyendaalseweg 135
City Nijmegen
ZIP/Postal code 6525 AJ
Country Netherlands
Platforms (1)
GPL1261 [Mouse430_2] Affymetrix Mouse Genome 430 2.0 Array
Samples (6)
GSM747404 C2C12 untreated day3 rep1
GSM747405 C2C12 untreated day3 rep2
GSM747406 C2C12 untreated day3 rep3
BioProject PRJNA143943

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Supplementary file Size Download File type/resource
GSE30192_RAW.tar 35.3 Mb (http)(custom) TAR (of CEL)
Raw data provided as supplementary file
Processed data included within Sample table

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