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Gene expression profiling of miR-regulated genes in proliferating C2C12

ABSTRACT: We used microarrays to detail the global programme of gene expression upon the over-expression of seven different differentiation-associated, E1A-regulated microRNAs.

ORGANISM(S): Mus musculus

PROVIDER: GSE26764 | GEO | 2012/09/01

SECONDARY ACCESSION(S): PRJNA136315

REPOSITORIES: GEO

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Gene expression profiling of miR-regulated genes in proliferating C2C12

Project description:We used microarrays to detail the global programme of gene expression upon the over-expression of seven different differentiation-associated, E1A-regulated microRNAs. seven different microRNAs (miR-1, miR-34a, miR-22, miR-365, miR-29a, miR-145 and Let-7b) and a negative control (AllStar, Qiagen) were over-expressed at 50nM for 36hrs in proliferating myoblasts. Three independent biological experiments were performed.

2012-09-01 | E-GEOD-26764 | biostudies-arrayexpress

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Project description:Proliferating C2C12 myoblasts were induced to differentiate into myotubes and then infected with adenovirus expressing E1A (Ad-E1A), which induces cell cycle re-entry and dedifferentiation. We analyzed the transcriptional profile of E1A infected C2C12-myotubes through the Affymetrix Mouse Genome 430 2.0 Array, searching for genes that were significantly regulated between two independent biological replicates at two different time points (24h and 36h after infection with Ad-E1A). In addition, we took advantage of the E1A mutant known as YH47/dl928 (hereafter referred as YH47), which bears two mutations in the pocket-binding region of E1A (Y48H, C124G) able to disrupt the interaction with Rb and its cognate proteins and to impair cell-cycle re-entry phenotype. YH47 mutant was used to identify the Rb independent transcriptional reprogramming of C2C12. C2C12 cells were differentiated in vitro to myotubes as previously described. Myotubes were, then, infected with an adenovirus carrying the 12S form of E1A (dl520), with the YH47 E1A mutant (dl928) or with a control adenovirus (CTR) expressing a deletion of essentially the entire E1A gene (dl312). Two different time points after infection were considered (24 hours and 36 hours) to evaluate changes in C2C12 cells expression profile. Technical (A or B) and biological replicates (EXP1 or EXP2) were done for each condition.

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