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)
Project description:Skeletal muscle C2C12 cells were treated with or without trichostatin A (TSA), which induces differentiation. Total RNA was extracted and profiled at 5 time points after induction of differentiation (0, 4h, 12h, 24h, 48h).
Project description:MicroRNAs (miRNAs) play a critical role in cells differentiation by targeting protein coding genes. The expression level of miRNA and mRNA between D0 and D4 differentiated C2C12 showed a great significance. It is closely related to myogenesis, and is helpful to understand function of miRNAs and disease related to muscle. We performed microarray and transcriptome profiling in differentiating C2C12 cells cells (at 0 and 4 days named D0 and D4, respectively) to detail the expression of mRNA and miRNAs during differentiation.
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.
Project description:Mice pancreatic cancer derived exosomes(KPC-exosomes) could induce skeletal muscle cells(C2C12) insulin resistance and exosomal microRNAs (miRNAs) may exert an important effect (Sci Rep. 2017;7(1):5384). The data set GSE95741 provided the differentially expressed micro RNAs. In this work, we aim to detect the differentially expressed genes and explore the intrinsic link.
Project description:Identification of product of proteolysis during C2C12 myoblast differentiation using subtiligase N-terminomics. Different cell populations collected during a time-course of differentiation (4 days) were used for N-terminal labeling in a forward degradomics approach (n=2). Day0 population= Myoblasts, Day1 populations= live cells and dead cells, Day4 populations= Myotubes and Reserve cells. Additionally, cleavages events generated by mouse caspase-3 at early stages of differentiation (Day 0 and 1) was evaluated using a reverse degradomics approach on myoblasts and live cells (n=2).
Project description:To investigate the importance of functional regulation of PGAM2 by SUMO conjugation in the context of myogenic differentiation. We performed RNAseq analysis of WT and sumoylation deficient mutant (K176R mutatant) C2C12 cells at different differentiation days, i.e. day 0 and day4, respectively (n=3/group).