Fig. 3.

PAK4 was a target gene of miR-9-5p in CRC cells. a The potential binding sites of miR-9-5p and PAK4 mRNA, as well as the sequences in potential binding sites of mutant-type plasmid. b and c Dual luciferase reporter assays were performed in HCT116 and SW1116 cells with vectors including the putative miR-9-5p target sites in the 3′-UTR of PAK4 mRNA (wild-type) and mutant. Data were normalized against Renilla or firefly luciferase activity. d Quantitative real-time PCR was used to determine the mRNA levels of PAK4 in HCT116 and SW1116 cells transfected with miR-9-5p mimics or miR-NC. e Western blot analysis was used to determine the protein levels of PAK4 in HCT116 and SW1116 cells transfected with miR-9-5p mimics or miR-NC. All data are expressed as the means ± SD. **p < 0.01, compared with the miR-NC group