Fig. 4

Hsa_circ_0003258 serves as a sponge for miR-653-5p in PC cells. A Schematic drawing showing the putative binding sites of miRNAs related to hsa_circ_0003258. B Lysates from DU145 cells subjected to biotinylated-hsa_circ_0003258 pull-down assay and the expression levels of hsa_circ_0003258 measured by qRT-PCR. C The expression of the top three candidate miRNAs predicted by CircInteractome database were quantified by qRT-PCR after biotinylated-hsa_circ_00032583 pull-down assay in DU145 cells. D A schema of hsa_circ_0003258 wild-type (WT) and mutant (Mut) luciferase reporter vectors. E Luciferase activity was tested in DU145 cells co-transfected with luciferase reporter containing hsa_circ_0003258 sequences with wild type and mutant binding site of miR-653-5p and the mimic of miR-653-5p or control. F left The relative expression of miR-653-5p detected by qRT-PCR after over expression or silencing of hsa_circ_0003258 in DU145 cells. F right The relative expression of hsa_circ_0003258 detected by qRT-PCR after the transfection of miR-653-5p mimic or inhibitor. G Venn diagram showing the predicted target genes of miR-653-5p. H The qRT-PCR showing the mRNA level change of the predicated targets after silencing of hsa_circ_0003258. I The qRT-PCR showing the mRNA level change of the predicated targets after over expression of hsa_circ_0003258. J Western blot showing protein levels of ARHGAP5 after overexpression or silencing of hsa_circ_0003258. ns: Not Significant; * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001