Role of miR-200c in Myogenic Differentiation Impairment via p66Shc: Implication in Skeletal Muscle Regeneration of Dystrophic mdx Mice

Figure 1

miR-200c overexpression in myoblasts inhibits skeletal muscle differentiation in vitro. C2C12 myoblasts were infected either with a lentivirus encoding miR-200c or with a control virus. After selection with puromycin, cells were plated and shifted to differentiation medium for 3 days. (a) Representative images of anti-MyHC staining (green). Nuclei were counterstained with DAPI (grey). Immunofluorescence with anti-MyHC antibody showed a decrease in myotubes in miR-200c-overexpressing cells compared to control. Scale bar: 200 μm. (b) Bar graphs representing differentiation index (percentage of MyHC-positive cells), fusion index (percentage of nuclei within a myotube), and number of nuclei per myotube. miR-200c overexpression decreased all these parameters ( independent experiments; ). (c) A representative Western blot using ZEB1, MyHC, myogenin, and antibodies showed that protein levels decreased upon miR-200c overexpression and MyoD expression was not affected. α-Tubulin (TUB) was used as loading control.

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