Figure 5.

Verification of ssc-miR-146 direct targets of porcine IRS1 by luciferase reporter assay. (A) Three 3′-UTR sequences containing normal, mutagenic, and deleted binding sites were inserted downstream of the luciferase reporter. Six nucleotides of IRS1 3′-UTR were mutated or deleted to disrupt the binding with miR-146b seed regions. (B) Schematic diagram showing dual-luciferase reporter constructs of porcine IRS1 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 significant differences between miR-146b mimic and NC groups transfected with vectors containing normal IRS1 3′UTR (n = 8, * p < 0.05).