Fig. 4

Overexpression of bta-miR-365-3p promoted primary cattle myoblast differentiation. a The expression level of miR-365-3p during PBMs differentiation after treatment with OPmiR-365-3p for differentiation six days. b The expression level of bta-miR-365-3p after treatment with OPmiR-365-3p in PBMs at four days. c The expression level of bta-miR-365-3p after treatment with its inhibitor for four days. d The mRNA expression of MYOD1 and MYOG in PBMs after treatment with OPmiR-365-3p for four days. e The protein expression of MYOG and MYOD1 under the treatment of OPmiR-365-3p, and the statistic relatively rate grayscale values of MYOG or MYOD1 compared withβ-actin with Image Lab software. f The mRNA expression of MYOD1 and MYOG in PBMs after treatment with the inhibitor of bta-miR-365-3p for four days. g The protein expression of MYOG and MYOD1 under the treatment of bta-miR-365-3p inhibitor, and the statistic the relatively rate grayscale values of MYOG and MYOD1 compared withβ-actin with Image Lab software. h MyHC (red)-positive myotubes were detected by immunofluorescence after transfection with OPmiR-365-3p, scale bars: 200 μm. i MyHC (red)-positive myotubes were detected by immunofluorescence after transfection with the inhibitor of bta-miR-365-3p. Scale bars: 200 μm. j The fusion indexes calculation, the number of nuclei in the myotube as a percentage of the total nucleus. The ‘Treat’ indicated that the groups were treated with OPmiR-365-3p or the inhibitor. The ‘Control’ indicated the groups of their negative control. Values are mean ± SD for three biological replicates. Note: MYOD1: Myogenic differentiation 1; MYOG: Myogenin. SD: standard deviation. *P < 0.05, ** P < 0.01, *** P < 0.001