Fig. 3

Circ_0004585 directly interacts with miR-1248 in anoikis -resistant PCa cells. A Schematic representation of the sites of the probe specific to the back-splice junction of circ_0004585. B, C Gel electrophoretic and qRT-PCR results showed that circ_0004585 could be specifically enriched by circ_0004585 probe. Relative level of circ_0004585 was normalized to input. GAPDH was used as negative control. D The relative expression levels of four miRNAs candidates were detected by qRT-PCR in PC-3 and DU145 cell lysates. E Circ_0004585 was enriched by biotinylated wild-type miR-1248 (Bio-1248-wt) or its mutant (Bio-1248-mut), and qRT-PCR was used to determine the relative circ_0004585 and GAPDH mRNA levels. F RNA FISH images showed the colocalization of circ_0004585 and miR-1248 in the cytoplasm of PC-3 cells (circ_0004585 probe was labeled with Cy3, locked nucleic acid miR-1248 was labeled with Dig, and the nuclei were stained with DAPI). Scale bar, 10 μm. Data are presented as the means ± SEM of three independent experiments (*P < 0.05; ** P < 0.01)