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Transcriptomic analysis of C2C12 myotubes after adenoviral expression of Crebrf

ABSTRACT: Transcriptomic analysis of C2C12 myotubes after adenoviral expression of Crebrf

PROVIDER: PRJNA947908 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR23949128.fastq.gz	Fastqsanger.gz
	SRR23949129.fastq.gz	Fastqsanger.gz
	SRR23949130.fastq.gz	Fastqsanger.gz
	SRR23949131.fastq.gz	Fastqsanger.gz
	SRR23949132.fastq.gz	Fastqsanger.gz

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Transcriptomic analysis of C2C12 myotubes after adenoviral expression of Crebrf

Project description:Purpose: identify global changes in gene expression in C2C12 myotubes caused by an increase in Crebrf. Methods: Triplicate replicates of differentiated C2C12 myotubes expressing either Gfp or Crebrf for 3 days after transduction. After assessing RNA quality with Agilent Bioanalyzer, mRNAs were enriched by poly-A pull-down. Then, sequencing libraries constructed with Illumina TruSeq RNA prep kit were sequenced using. We multiplexed samples in each lane, which yields targeted number of single-end 75 bp reads for each sample, as a fraction of 180 million reads for the whole lane. Sequence reads were mapped back to the Drosophila genome (flybase genome annotation version r6.30) using STAR. With the uniquely mapped reads, we quantified gene expression levels using Cufflinks (FPKM values). Next, differentially expressed genes between experimental and control data were analyzed with DESeq2. Results: Gene set enrichment analysis reveals oxidative phosphorylation and interferon response as the top up- and down-regulated gene sets, respectively, upon Crebrf expression. The glycolysis gene set is also reduced. These gene expression findings parallel our metabolic observations in Seahorse analysis. Conclusions: Our study indicates that CREBRF is a strong regulator of muscle metabolism in C2C12 myotubes

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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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circRNA expression in C2C12 myoblasts and myotubes

Project description:circRNA expression in C2C12 myoblasts and myotubes

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Gene expression profile of E1A infected C2C12 myotubes

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Project description:We analyzed the change of H3K27Ac globally in response to microgravity in C2C12 mouse myotubes.

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