Fig. 4

Overexpression of miR-542-3p inhibits myoblast differentiation. A The mRNA level of MYOG, MYOD1, MYHC, MYF5, and MYF6 at day 3 after transfection of miR-542-3p mimic and control groups. B, C The western blotting bands of differentiation markers in the miR-542-3p mimic and control groups (B) and their quantitative analysis (C) by image J software. The membrane was cleaved before hybridization with the antibody, and all protein bands (MYOG, MYOD1, and β-actin) in Fig. 4B were from the same samples. 4 biological replicates were detected, 3 of which were shown in the Fig. 4B and full-length blots were presented in Supplementary Fig. S4. D Anti-Myosin heavy chain (MYHC) immunofluorescence staining after the transfection of miR-542-3p mimic, scale bar: 100 μm. E, F, G MYHC immunofluorescence intensity analysis (E), differentiation index (F), myotube fusion index (G) by image J software. n = 3, * P < 0.05, ** P < 0.01