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Genomic mapping of Six1 binding sites during skeletal myoblast differentiation

ABSTRACT: In this study, the C2C12 cell line, a model used to study myogenesis and regeneration, was allowed to differentiate from myoblast precursor cells to myotubes. Cells were harvested at 3 different timepoints to perform ChIP-on-Chip of Six1, which is a key muscle regulator. We identified global loci bound by Six1 during skeletal myoblast differentiation.

ORGANISM(S): Mus musculus

PROVIDER: GSE20150 | GEO | 2010/07/05

SECONDARY ACCESSION(S): PRJNA125575

REPOSITORIES: GEO

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Genomic mapping of Six1 binding sites during skeletal myoblast differentiation

Project description:In this study, the C2C12 cell line, a model used to study myogenesis and regeneration, was allowed to differentiate from myoblast precursor cells to myotubes. Cells were harvested at 3 different timepoints to perform ChIP-on-Chip of Six1, which is a key muscle regulator. We identified global loci bound by Six1 during skeletal myoblast differentiation. C2C12 Myoblasts were allowed to differentiate into myotubes. Cells at three timepoints were harvested for ChIP-on-Chip, including myoblasts stage, 24h after differentiation and myotubes (96h after differentiation). Myotubes were detached from the undifferentiated myoblast reserve cells using diluted trypsin. 3 independent biological replicates were used for each time point experiment. A microarray set counts 3 arrays (Custom Arrays A, B and C) for a total of approximately 2.9 million probes.

2010-07-05 | E-GEOD-20150 | biostudies-arrayexpress

Gene Expression Profiling of C2C12 Myoblast Differentiation

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Quantitative Myogenesis C2C12 Proteome with Dimethyl-Labelling

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C2C12 murine myoblast cell line and 48 h 10 uM BIO treated C2C12 cellulate

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Novel RNA-binding activity of MYF5 enhances Ccnd1/Cyclin D1 mRNA translation during myogenesis

Project description:Skeletal muscle contains long multinucleated and contractile structures known as muscle fibers, which arise from the fusion of myoblasts into nucleated myotubes during myogenesis. The myogenic regulatory factor (MRF) MYF5 is the earliest to be expressed during myogenesis and functions as a transcription factor in muscle progenitor cells (satellite cells) and myocytes. In mouse C2C12 myocytes, MYF5 is implicated in the initial steps of myoblast differentiation into myotubes. Ribonucleoprotein immunoprecipitation (RIP) analysis showed that MYF5 bound a subset of myoblast mRNAs; prominent among them was Ccnd1 mRNA, which encodes the key cell cycle regulator CCND1 (Cyclin D1). Biotin-RNA pulldown, UV-crosslinking, and gel shift experiments indicated that MYF5 was capable of binding the 3' untranslated region (UTR) and the coding region (CR) of Ccnd1 mRNA. MYF5 silencing in proliferating growing myoblasts revealed that and MYF5 promoted CCND1 translation, and it also modestly increased transcription of Ccnd1 mRNA. Importantly, silencing MYF5 reduced myoblast growth as well as differentiation of myoblasts into myotubes, while overexpressing MYF5 in C2C12 cells upregulated CCND1 expression. We propose that MYF5 enhances early myogenesis in part by coordinately elevating Ccnd1 transcription and Ccnd1 mRNA translation. Four replicates were utilized from either Control (IgG) or MYF5-immunoprecipitated RNA samples from C2C12 cells growing in either growth medium (GM) or differentiation medium (DM) for a total of sixteen samples.

2016-08-08 | E-GEOD-73704 | biostudies-arrayexpress

C2C12 murine myoblast cell line and 48 h 10 uM BIO treated C2C12 cellulate

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Gene expression analysis of differentiating skeletal muscle myoblast cell line overexpressing MyoD

Project description:We tested the hypothesis that ectopic expression of the MyoD-dependent gene program in differentiating myoblasts and myotubes compromises the integrity of the plasma membrane/sarcolemma by using a doxycycline-inducible system to overexpress MyoD in C2C12 myoblasts during differentiation. The overarching goals of this analysis pipeline were to (a) determine changes in transcriptional profiles upon sustained overexpression of MyoD during and upon differentiation; and, (b) utilize these profiles to identify potential drivers of sarcolemmal fragility that exacerbate myofiber damage and loss in dystrophic muscle. We used microarrays to generate information about transcriptional profiles in differentiating myoblasts and myotubes overexpressing the myogenic transcription factor myoblast determination protein 1 (MyoD1).

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Gene Expression Profiling of siRNA Knock-Down of Six1, Six4 and Myogenin in C2C12

Project description:Six1, Six4 and Myogenin are transcription factors that are known to be required for skeletal myogenesis. Currently, very little is known about the genes targeted by Six1 and Six4. Gene expression profiling when one or both transcription factors were knock-down by siRNA was performed to identify genes affected by their absence. We also hypothesized that Six1 and Six4 can work in cooperation with the myogenic regulatory factor (MRFs) family of transcription factors, such as Myogenin. Therefore, we performed the same type of experiment where the myogenin was knocked-down by siRNA to identify genes that are possibly regulated by the Six1 or Six4 in conjunction with Myogenin. C2C12 Myoblasts were transfected with siRNA against Six1, Six4, Six1 with Six4, Myogenin, or control 24h before start of differentiation. The cells were allowed to differentiate in differentiation medium for 24h and were harvested for gene expression profiling. Four replicates per siRNA were performed.

2010-07-30 | E-GEOD-19967 | biostudies-arrayexpress

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