Project description:We used microarrays to characterize the global changes in gene expression in C2C12 cells due to siRNA knockdown of long non-coding RNA H19 Control siRNA or siRNA specific for mouse H19 were transfected into day1 differentiating C2C12 myoblasts in triplicates. 40 H later total RNAs were isolated and subjected with microarray analysis.
Project description:To determine the lncRNA expression profile in C2C12 myoblasts and myotubes, we used mouse lncRNA microarray from Arraystar to examine the expression of lncRNAs in C2C12 myoblasts and myotubes.
Project description:To determine the circRNA expression profile in C2C12 myoblasts and myotubes, we used mouse circRNA microarray from Arraystar to examine the expression of circRNAs in C2C12 myoblasts and myotubes.
Project description:C2C12 myoblasts differentiation is a precise controlled process. Splicing of many genes changes during the differentiation process. Some muscle-specific splicing isoforms play important role in myogenesis. We used microarrays to detail the global programme of gene expression underlying defect of lrrfip1a, a muscle specific splicing isoform of Lrrfip1. Differentiated C2C12 myoblasts shLUCI or shLrrfip1a were collected and extracted RNA, then hybridizated on Affymetrix. We sought to obtain the different expressed genes between shLUCI and shLrrfip1a samples.
Project description:We report that differentiation of the C2C12 myoblast line on patterened hydrogels results in the increased expression of muscle sarcomere genes and more closely models expression changes that happen in in vivo muscle differentiation than do C2C12 myoblasts cultured on unpatterened substrates.
Project description:Differentiation of muscle tissue is regulated by a complex network of transcription factors. The MEF2 family of transcription factors are important players in muscle development and differentiation. We knocked down expression of MEF2 isoforms in a cell culture model of skeletal muscle differentiation and assessed global expression pattern changes between MEF2 knockdowns, and with a negative control. C2C12 myoblasts were transduced with shRNA viral vectors to knockdown expression of individual MEF2 proteins. Cells were then induced to differentiate and were harvested 3 days post-differentiation for Affymetrix global gene expression analysis. Each array was pooled RNA from two separate transductions, and each treatment group was analyzed in triplicate.
Project description:Analysis of differentiating LSD1-KD C2C12 myoblasts. We found LSD1 is an important regulator of oxidative phenotypes in skeletal muscle cells. Results provide insight into the molecular mechanisms underlying roles of LSD1 in myocytes.
Project description:C2C12 myoblasts differentiation is a precise controlled process. Splicing of many genes changes during the differentiation process. Some muscle-specific splicing isoforms play important role in myogenesis. We used microarrays to detail the global programme of gene expression underlying defect of lrrfip1a, a muscle specific splicing isoform of Lrrfip1.
Project description:We show the application of 5mC antibody-based methylated DNA immunoprecipitation followed sequencing technology for high-through profiling of DNA methylation in mouse C2C12 myoblasts and myotubes. By analyzing the methylation status of immunoprecipitated DNA fragments, we generated genome-wide DNA methyaltion maps in mouse C2C12 myoblasts and myotubes. We find that DNA methylation levels in myoblasts at rDNA promter and coding regions are higher than that in myotubes but not changed in intergenic regions.
Project description:In urodele amphibians, limb regeneration involves the dedifferentiation of muscle myotubes into single cells that may acquire pluripotent potential. We have employed small molecules (myoseverin and BIO) to attempt to reproduce this behavior in mammalian muscle culture. C2C12 myotubes derived from the C2C12 myoblast cell line were induced to undergo cellularization by myoseverin treatment, which destabilizes tubulin filaments. The GSK-3 inhibitor, BIO, was then used to induce dedifferentiation. Induce neuron formation; the cells were incubated with 250 nM reversine for 48 h, and neural induction media (DMEM/F12 supplemented with N2 (Invitrogen)) and 1.5 uM all-trans retinoic acid for 7 days. C2C12 murine myoblast cell line and 48 h 10 uM BIO treated C2C12 cellulate (derived by 20 M myoseverin treatment for 48 h) C2C12 myoblasts were differentiated into myotubes with 2%horse serum in DMEM for 8 days (from 2-4 d, 10 uM AraC treatment was also used to kill any remaining myoblasts). Next, myotubes were cellularized by 20 uM myoseverin treatment for 48 h. 24 h after myoseverin treatment, myotubes were treated with 10 uM BIO for 2d.
