Figure 4. NEDD9 was a direct target of miR-1252-5p in PAC cells. (A) miR-1252-5p and its putative binding sequences in the 3′-UTR of NEDD9. (B–E) qRT-PCR and western blot analysis of NEDD9 mRNA and protein expression after transfection with corresponding miRNA vectors in PAC cells. (F) Western blot assays of NEDD9 protein expression in human PAC tissues with high- and low-miR-1252-5p expressing. (G) An inverse correlation between miR-1252-5p and NEDD9 mRNA levels was observed in human PAC tissues (n = 18). (H, I) Luciferase activity was detected after co-transfection with corresponding miRNA vectors and luciferase vectors in PAC cells. NEDD9, neural precursor cell expressed, developmentally downregulated 9; PAC, pancreatic cancer; WT, wild-type; MUT, mutant-type; UTR, untranslated region. **P < 0.01; ***, P < 0.001, ns, not significant.