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

Figure 2

miR-200c overexpression in myotubes inhibits skeletal muscle differentiation in vitro. C2C12 myoblasts were shifted to differentiation medium for 24 hrs; then, cells were infected either with a lentivirus encoding miR-200c or with a control virus. Afterwards, cells were selected with puromycin in differentiation medium for 3 days. (a) Representative-phase contrast images of C2C12 myoblasts prior to infection (upper panels) and after infection (lower panels). (b) Representative images of anti-MyHC staining (green). Nuclei were counterstained with DAPI (grey). Scale bar: 200 μm. (c) Bar graphs representing differentiation index, fusion index, and number of nuclei per myotube. miR-200c overexpression decreased all these parameters ( independent experiments; ). (d) A representative Western blot using ZEB1, MyHC, myogenin, and MyoD antibodies showed that protein levels decreased upon miR-200c overexpression. α-Tubulin (TUB) was used as loading control.

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