Figure 4

RKIP is a direct target of miR-27a. (A) The predicted miR-27a binding site within RKIP 3′UTR and its mutated version by site mutagenesis are as shown. (B) Variable RKIP expression in A549 and A549/CDDP was obtained by western blot. (C) A549 cells were transfected with NC or miR-27a mimics, A549/CDDP cells were transfected with anti-NC or miR-27a inhibitors respectively. Western blotting was used to detect RKIP expression; β-actin was used as an internal control. (D) Luciferase assay was performed in A549 cells that were co-transfected with miRNA mimics and reporter vectors carrying RKIP 3′ UTR with wild type versus mutated miR-27a response element. Data are means of three separated experiments ± SD; *P < 0.01.