Figure 5.

Mechanism dissection on how miR-616-5p alters NR2C2 expression: via binding to the 3′UTR of NR2C2. (A) RNA binding protein immunoprecipitation assay for miRNAs using IgG or AGO2 antibody in T24 cells and UMUC3 cells. The qRT-PCR assay was used to detect NR2C2 expression in each group. (B) Sequence alignment of NR2C2 3′UTR with wild-type versus mutant potential miR-616-5p targeting sites. The wild-type and mutant psiCHECK2- NR2C2 3′UTR reporter constructs. (C) Luciferase reporter activity after transfection of the wild-type or mutant NR2C2 3′UTR reporter construct in T24 cells transduced with pLKO and miR-616-5p inhibitor and UMUC3 cells transduced with pLKO and oemiR-616-5p. All quantification data are presented as mean ± SD, and significant differences are indicated by and ****P < 0.0001 and ns, not significant compared to the controls.