Fig. 5

NLK functioned as the target of miR-665 in CRC cells.AExpression of potential target genes in CRC cells transfected with OE-circ_0009092, SH- circ_0009092. (B) qRT-PCR analysis of potential target genes in CRC cells transfected with miR-665 mimic, mimic NC, miR-665 inhibitor, inhibitor NC.CNLK protein expression was determined by Western blot in HCT116 and Lovo cells transfected with OE-circ_0009092, SH- circ_0009092.DNLK protein expression was determined by Western blot in HCT116 and Lovo cells transfected with miR-665 mimic, mimic NC, miR-665 inhibitor, inhibitor NC.EThe expression of NLK in 80 paired CRC tissues was detected by qRT-PCR.FThe correlation between NLK and miR-665 expression in 80 paired CRC tissues.GSchematic representation of potential binding sites between miR-665 and NLK.HRIP assay was conducted for detecting the binding of miR-665 and NLK by using the Ago2 antibody in CRC cells transfected with miR-665 mimic, mimic NC.IRelative luciferase activities were determined in CRC cells transfected with NLK 3′-UTR WT or MUT luciferase reporter vectors and miR-665 mimic or mimic NC.JWestern blot showed the protein changes of E-cadherin, Vimentin, and NLK in control, OE-circ_0009092, miR-665 mimic, and OE-circ_0009092+miR-665 mimic -treated HCT-116 cells.KWestern blot showed the protein changes of Wnt/β-catenin signaling pathway related proteins in control, OE-circ_0009092, miR-665 mimic, and OE-circ_0009092+miR-665 mimic-treated HCT-116 cells.LWestern blot showed the expression changes of E-cadherin, Vimentin, and NLK in Lovo cells by treatment of SH-circ_0009092, miR-665 inhibitor, and SH-circ_0009092+miR-665 inhibitor.MWestern blot showed the protein changes of Wnt/β-catenin signaling pathway related proteins in SH-circ_0009092, miR-665 inhibitor, and SH-circ_0009092+miR-665 inhibitor-treated Lovo cells. Statistical analysis between two groups was performed using two-tailed t-test. One-way ANOVA statistical tests were adopted for more than two groups. Data are the means ± SD of three experiments. *P< 0.05, **P< 0.01, ***P< 0.001