Fig. 5

miR-29c-3p directly binds to circ-001971 and VEGFA (a-b) Two different types of luciferase reporter vectors were constructed: wild-type circ-001971 and VEGFA 3’UTR, and mutant-type circ-001971 and VEGFA 3’UTR; mutant-type vectors contained a 4 bp mutation in the predicted miR-29c-3p binding site. c-d The above vectors were cotransfected into HEK293 cells with miR-29c-3p mimics or miR-29c-3p inhibitor and examined for luciferase activity. e RIP assays were performed to confirm the binding of miR-29c-3p to circ-001971 and VEGFA using an AGO2 antibody. The levels of miR-29c-3p, circ-001971 and VEGFA in precipitated AGO2 proteins were examined using real-time PCR. f Endogenous circ-001971 and VEGFA 3’UTR pull-down by AGO2 upon overexpression of miR-29c-3p was determined using RIP assays. The data are presented as the mean ± SD of three independent experiments. **P < 0.01, compared to mimics-NC, inhibitor-NC, IgG group