Fig. 6

CircPPAP2B functions as a ceRNA to directly bind to miR-182-5p and upregulated CYP1B1 A Bioinformatics tools (circBank, ENCORI, and CircInteractome) were used to identify candidate miRNAs that potentially interact with circPPAP2B. B Transwell assays were performed to explore the role of miR-182-5p mimic or miR-182-5p inhibitor on the invasive ability of ccRCC cells. C Biotin-labeled RNA pulldown assays were performed to confirm the direct interaction between circPPAP2B and miR-182-5p. D Dual-luciferase reporter assay was performed to validate the direct binding and binding site between circPPAP2B and miR-182-5p. E Transwell assays were performed to explore the role of miR-182-5p mimic or miR-182-5p inhibitor on the circPPAP2B-mediated invasion of ccRCC cells. F High-throughput RNA sequencing to analyze the RNA regulated by circPPAP2B in ccRCC cells. G Western blotting assays to investigate the effect of miR-182-5p on circPPAP2B-mediated CYP1B1 expression. H Biotin-labeled RNA pulldown assay was performed to validate the direct binding between miR-182-5p and CYP1B1 mRNA. I Dual-luciferase reporter assay was performed to validate the direct binding and binding site between miR-182-5p and 3’UTR of CYP1B1 mRNA. J Transwell assay was performed to explore the role of CYP1B1 in the invasion of ccRCC cells. Data are represented as mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001 vs. WT group