Figure 3.

STK11 is a direct target of miR-424 and is positively regulated by miR-424 in bovine preadipocytes. (A) (Top) Schematic representation of the sequence of mature miR-424. The putative binding sites were located in three prime untranslated region (3' UTR) of the bovine STK11 gene, and the point mutation generated is underlined. (Bottom) Relative luciferase activity was measured in HEK293A cells co-transfected with the psi-Check2 empty vector (EV), STK11 3' UTR-WT, or STK11 3' UTR-MUT reporter plasmids along with NC mimics or miR-424 mimics. Renilla luciferase activity was normalized to firefly luciferase. (B) The messenger RNA (mRNA) and protein expression of STK11 in bovine preadipocytes were examined 48 h post-transfection with miR-424 mimics and inhibitor. The quantitative representation of the gray ratio of STK11/β-actin protein is displayed numerically under the strip. Results are shown as the mean ± SEM and the data are representative of three biological and technical replicates. *p <  0.05, **p <  0.01, n.s., not significant.