Figure 3.

Overexpression of miR-744 Inhibits Differentiation of Bovine Primary Myoblasts

(A–C) Myoblasts were treated with pcDNA3.1 or pcDNA3.1-miR-744. (A and B) The mRNA and protein expression of cell differentiation markers, MyoG, MyoD, and MyHC, was measured by real-time qPCR (A) and western blotting (B). (C) Cell differentiation was also detected by immunofluorescence; the scale bars represent 200 μm. (D) The efficiency of miR-744 knockdown was evaluated by qPCR. (E) Real-time qPCR analysis of the differentiation-related genes. (F) Western blot analysis of MyOG, MyOD, and MyHC protein levels with β-actin treatment. (G) 293T cells were co-transfected with pcDNA3.1-miR-744 and psi-CaMKIIδ (or) psi-Wnt5a-3′ UTR-W/Mut. (H and I) The mRNA and protein expression of CaMKIIδ and Wnt5a was measured via real-time qPCR (H) and western blotting (I). Values are mean ± SEM for three biological replicates; *p < 0.05; **p < 0.01; ***p < 0.001; NS, no significant differences.