Figure 4.

MiR-142-3p directly targets HMGB1 mRNA and inhibits its expression. A. The putative miR-142-3p-binding sites in the 3’-UTR of HMGB1 mRNA was shown. Mutation was generated on the HMGB1 3’-UTR sequence in the complementary site for the seed region of miR-142-3p, as shown. B. The wild type (HMGB1 3’-UTR-WT) or mutant (HMGB1 3’-UTR-mut) reporter plasmids was co-transfected into NCI-H23 and NCI-H838 with miR-142-3p or miR-control. The normalized luciferase activity in the control group was set as relative luciferase activity. C. The expression of HMGB1 mRNA was analyzed by real time PCR assay. ACTIN was used as an internal control. D, E. The expression of HMGB1 protein was analyzed by western blot assay. actin was used as an internal control. All experiments were at least repeated in triplicate with similar results.