Figure 5.

MiR-149-5p directly targeted WT1 in HCC. (a) The predicted binding sites between miR-149-5p and the 3ʹ-UTR of WT1 or MUT WT1. (b) The interactions of miR-149-5p with wild-type WT1 or mutated WT1 were verified by dual luciferase reporter assay (*P < 0.05 vs. miR-NC group). (c) Cells were transfected with miR-149-5p mimics or miR-NC (negative control). The amount of miR-149-5p and WT1 in HEK-293 T cells was detected after Ago2 RNA immunoprecipitation (RIP) (*P < 0.05 vs. IgG group). (d) qRT-PCR analysis of the expression of WT1 in normal and HCC tissues (n = 35, *p < 0.05 versus Normal). (e) Immuno-blotting analysis of WT1 expression after transfection with miR-149-5p mimics or WT1 pDNA as indicated, miR-NC and pcDNA were used as the negative control (*P < 0.05 versus miR-NC or “miR-149-5p+pcDNA”). (f) The correlation between miR-149-5p and WT1 mRNA expression. Data were presented as mean±SEM (n ≥ 3)