Fig. 2

Overexpression of miR-542-3p inhibits myoblast proliferation. A The expression level of miR-542-3p in C2C12 myoblasts at 48 h after its mimic transfection was determined by RT-qPCR. B The mRNA level of CCND1, CCND2, CCNB1, CDK2, CDK4, and P53 in the miR-542-3p mimic and control groups. C, D The western blotting bands of proliferation markers in the miR-542-3p mimic and control groups (C) and their quantitative analysis (D) by image J software. The membrane was cleaved before hybridization with the antibody, and all protein bands (CCND1, CCNB1, and β-actin) in Fig. 2C were from the same samples. 4 biological replicates were detected, 3 of which were shown in the Fig. 2C and full-length blots were presented in Supplementary Fig. S2. E, F The images of 5-ethynyl-20-deoxyuridine (EdU) staining (E) and percentage of the EdU positive cells (F), scale bar: 100 μm. G Cell viability was measured by cell counting kit-8 assay (CCK-8). n = 3, * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001