Fig. 3

CircSTX6 directly interacts with miR-515-3p in BCa. A The potential binding miRNAs with circSTX6 were predicted by the databases of CircInteractome and CircBank. B Schematic diagram showed the putative binding sites of three miRNA candidates associated with circSTX6. C The efficiency of circSTX6 probe were tested by RNA pulled down and qRT-PCR. D Relative levels of three miRNAs in EJ and UMUC3 lysates pulled down by circSTX6 probe or oligo probe. E RIP assays were performed to detect the enrichment of circSTX6 and miR-515-3p upon AGO2 antibody in BCa cells. F Schematic of circSTX6 wild-type (wt) and mutant (mut) luciferase reporter vectors. G Luciferase reporter assays revealed the relative luciferase activity of wild type or mutant circSTX6 in the miR-515-3p mimics or NC group. H The RNAs captured by biotinylated wild-type or mutant miR-515-3p BCa cells were quantified by qRT-PCR. I Co-localization of circSTX6 and miR-515-3p in BCa cells was detected by FISH assay. Scale bar, 10 μm. J Transwell assays showed the metastasis in BCa cells stably transfected with scramble or sh-circ#1, and those co-transfected with anti-miR-NC or anti miR-515-3p. Scale bar, 100 μm. The data are presented as the means ± S.D. of at least three independent experiments. *P < 0.05. P values are calculated by Student’s t test in C-E, G and H and one-way ANOVA in J