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. 2013 Dec 27;15(1):296–308. doi: 10.3390/ijms15010296

Figure 3.

Figure 3.

Overexpression of miR-199a-3p inhibited C2C12 myogenic differentiation. C2C12 cells were transfected with miR-199a-3p Mimics (or negative control miRNA) and grown in growth medium (GM) for 12 h, then induced to myogenic differentiation with differentiation medium (DM). At the 96 h of differentiation: (A) Immunofluorescence of muscle myosin heavy chain (MyHC) in C2C12 myotubes; (B) The over-expression efficiency was measured by real-time qPCR; (C) The fusion index was counted by MyHC-positive cells to total nuclei (Hoechest-positive cells); (D) Myotubes sizes were determined by myonuclei counts in myotubes transfected with miR-199a-3p mimic or negative control miRNA; (E) Relative mRNA levels of MyoD and Myf5 were detected by real-time PCR at the 24 h of differentiation; (F) Relative mRNA levels of MyoG, MyHC, MURF1 and Atrogin1 at the 96 h of differentiation; (G) Western Blot detected the protein expression of myogenic markers MyoG, and MyHC at the 96 h of differentiation; and (H) The relative protein levels in (G) were analyses using image J. Results are presented as mean ± S.E.M. n = 3. * p < 0.05, ** p < 0.01.