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

Figure 3

Anti-miR-200c treatment enhances skeletal muscle differentiation in vitro. C2C12 myoblasts were infected either with a lentivirus encoding anti-miR-200c or with a control virus. After selection with puromycin, cells were plated and shifted to differentiation medium for the times indicated in the figure. (a) Representative images of anti-MyHC staining (green). Nuclei were counterstained with DAPI (blue). Immunofluorescence with anti-MyHC antibody showed an increase in myotubes in anti-miR-200c-overexpressing cells compared to control at 3 days in DM. Scale bar: 100 μm. (b) Bar graphs representing differentiation index, fusion index, and number of nuclei per myotube. Anti-miR-200c overexpression increased all these parameters ( independent experiments; ). (c) A representative Western blot using MyHC, myogenin, and MyoD antibodies showed that all these protein levels increased upon anti-miR-200c overexpression. α-Tubulin (TUB) was used as loading control.

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