Fig. 4

Circ_0004585 binds miR-1248 to upregulate the transcriptional activity of TM9SF4 and promote cell invasion in anoikis -resistant PCa cells. A Representative images (left panel) and quantification (right panel) results of transwell migration assay for PC-3 and DU145 transfected with miR-1248 mimics alone or cotransfected with circ_0004585 (Scale bar, 100 μm). B Representative images and quantification results of wound-healing assay for PC-3 and DU145 transfected with miR-1248 mimics alone or cotransfected with circ_0004585 (Scale bar, 100 μm). C Apoptosis was assessed by flow cytometry assay in PC-3 and DU145 transfected with miR-1248 mimics alone or cotransfected with circ_0004585. D The Venn diagram shows TM9SF4 is commonly predicted by TargetScan, miRDB and PicTarBase. E Schematic graph illustrated the mutation of potential binding site between miR-1248 and the 3′-UTR regions of TM9SF4. F Luciferase activity in PCa cells cotransfected with a luciferase reporter containing either TM9SF4 3′-UTR-wt or TM9SF4 3′-UTR-mut and miR-1248 mimics or inhibitors. G The protein levels of TM9SF4 detected by western-blotting in miR-1248 overexpressing and knockdown cells. Bar graphs show the statistical analysis of three independent experiments (* P < 0.05; ** P < 0.01)