Fig. 7

CircSLC6A6 functions as an efficient miR-1265 sponge, and alters the expression of C2CD4A through miR-1265 a Representative FISH images showed the co-location between circSLC6A6 and miR-1265 in RKO and HCT8 cells. b Correlations between circSLC6A6 with miR-1265 and C2CD4A mRNA expression were performed by Pearson’s correlation analysis in CRC tissue samples (n = 63). c qRT-PCR was applied to determine the effect of altered expression of circSLC6A6 on miR-1265 expression in RKO and HCT8 cells. d Putative binging sites between miR-1265 and circSLC6A6 were predicted by CircInteractome, and the schematic of circSLC6A6 wild-type (WT) and mutant (Mut) luciferase reporter vectors. e Luciferase activity of circSLC6A6-WT, circSLC6A6-Mut in RKO and HCT8 after co-transfection with miR-1265 cells mimics, inhibitor or miRNA control. f Anti-Ago2 RIP was performed in RKO cells transfected with miR-1265 mimics or miR-NC, then circSLC6A6 expression detected by agarose gel electrophoresis and qRT-PCR. g In RKO cells, endogenous miR-1265 was pulled down and enriched with circSLC6A6 probe, then the enrichment of miR-1265 was detected by qRT-PCR. Biotin-miR-1265 captured endogenous circSLC6A6 in the cell complex and was compared with biotin-NC then the enrichment of circSLC6A6 was detected by using qRT-PCR. Calculation of experiment results were followed by the ratio of pull-down to input. h Proposed model for circSLC6A6/miR-1265/C2CD4A/p53 axis regulatory network, circSLC6A6 as a ceRNA for miR-1265 to regulate C2CD4A and p53 expression in CRC. All data are presented as the mean ± SEM. (ns showed no significance. *P < 0.05, **P < 0.01, ***P < 0.001)