Fig. 2.

MUC4 is a direct target of miR-150. (A) Schematic representation of firefly luciferase reporter construct containing MUC4 3′ UTR with either wild-type (WT) or mutant (MUT) miR-150 target site. MUT-3′ UTR construct carries three nucleotides (74–76) variation in the seed matching region of the target site to disrupt binding of miR-150. (B) Luciferase reporter assay to examine the miR-150-mediated control of gene expression. HPAF, Panc10.05 and Colo357 cells (0.5 × 10⁶ cells per well) were transiently cotransfected for 24 h with reporter plasmids (200 ng, WT or MUT) and 100 nM of miR-150 or miR-NC mimic. Subsequently, protein lysates were made and luciferase (Firefly; test and Renilla, transfection efficiency control) activity assessed using a dual-luciferase assay system. Data are presented as normalized fold change in luciferase activity (mean ± SD; n = 3, *P < 0.05).