Fig. 3

CircBNC2 functions as a sponge for miR-4298. A The Venn diagram showing the predicted overlapping potential target miRNAs of circBNC2 by TargetScan, circbank and miRDB. B Correlation analysis showing the mRNA expression between circBNC2 and miR-4298 in 78 PCa patient samples. C The co-localization of circBNC2 and miR-4298 in DU145 and PC3 cells was analyzed by FISH assay. The circBNC2 probe was labeled with Cy3 (red), miR-4298 probes were labeled with FAM (green), and nuclei were stained with DAPI (blue). Scale bar = 10 μm. D The schematic illustration for construction of luciferase reporter plasmids, wild-type (WT) and mutant (Mut)-circBNC2, as indicated (left panel). The luciferase activity was measured in DU145 and PC3 cells which co-transfected with of the luciferase reporter plasmid WT/Mut-circBNC2 and miR-4298 mimic/NC mimic, respectively (right panel). E Fold enrichment of circBNC2 and miR-4298 in PC3 and DU145 cells by the AGO2-RIP assay. F and G RT-qPCR analysis for the expression of miR-4298 in OE-circBNC2 (F) or sh-circBNC2 (G) DU145 and PC3 cells. H - J In OE-Ctrl + NC mimic-, OE-Ctrl + miR-4298 mimic-, OE-circBNC2 + NC mimic- or OE-circBNC2 + miR-4298 mimic- treated DU145 and PC3 cells, cell proliferation, invasion and migration were analyzed by the CCK-8 assay (H), transwell assay (I) (Scale bar = 50 μm) and wound healing assay (J) (Scale bar = 200 μm), respectively. K - M The representative images of subcutaneous xenograft tumors (n = 6/group) (K), tumor growth curves (L), and tumor weight (M) at the endpoint from control and circBNC2 overexpression groups co-treated with NC agomir or miR-4298 agomir. ns, no significance; *P < 0.05; **P < 0.01; ***P < 0.001