miR-130b targets on the 3′-UTR of MyHC IIx.
A, putative binding site of miR-130b on 3′-UTR of MyHC IIx was predicted by RNA-hybrid tool. B, enrichment of MyHC IIx mRNA immunoprecipitated by biotin-labeled miR-130b or miR-130b–Mut probes was determined RNA hybrid qRT-PCR (n = 3). C, statistical results of dual-luciferase reporter assay (n = 5). D, levels of MyHC IIx mRNA in different skeletal muscles of 180-day-old pigs was estimated through qRT-PCR (n = 5). E and F, expression of MyHC IIx at protein levels in different skeletal muscles of 180-day-old pigs was detected by Western blotting (n = 5). G, reduced MyHC IIx was restored by miR-130b inhibitors at mRNA levels (n = 3). H and I, repressed MyHC IIx was rescued by miR-130b inhibitors at the protein level (n = 3). NC, negative control. *, p < 0.05; **, p < 0.01.