Figure 4.

ssc-miR-146b not directly targets porcine GLUT4 by luciferase reporter assay. (A) Three 3′-UTR sequences containing normal, mutagenic, and deleted binding sites were inserted downstream of the luciferase reporter. Seven nucleotides of GLUT4 3′-UTR were mutated and deleted to disrupt the binding with miR-146b seed regions; (B) Schematic diagram showing dual-luciferase reporter constructs of pig GLUT4 3′-UTR with putative miR-146b binding site; (C) Constructed vectors were transfected into Hela cells with miR-146 mimics or NC. The luciferase assay results revealed no significant differences betweenmiR-146b mimics and NC transfected with vectors containing normal GLUT4 3′-UTR, mutant GLUT4 3′-UTR and the deletion (n = 8).