Fig. 6

MiR-599 directly interacted with ONECUT2. (A) ONECUT2 was predicted to be the target gene of miR-599. (B-E) The combination between miR-599 and ONECUT2 was assessed by RIP and RNA pull down assays. (F) The binding sites between ONECUT2 and miR-599. (G and H) The combination between miR-599 and ONECUT2 was analyzed by dual-luciferase reporter assay. (I and G) The mRNA and protein levels of ONECUT2 in SW620 and DLD-1 cells transfected with miR-NC or miR-599 were measured by qRT-PCR assay or western blot assay. (K and L) The mRNA and protein levels of ONECUT2 in SW620 and DLD-1 cells transfected with sh-NC + anti-NC, sh-circ_0084615 + anti-NC or sh-circ_0084615 + anti-miR-599 were measured by qRT-PCR assay or western blot assay. (M and N) The expression of ONECUT2 in CRC tissues and normal tissues was examined with qRT-PCR and IHC. (O and P) The mRNA and protein levels of ONECUT2 in FHC, SW620, DLD1, SW480 and HT-29 cells were measured by qRT-PCR assay or western blot assay. *P < 0.05, **P < 0.01, ***P < 0.001