Fig. 1

The mechanism of regulatory relationships among EZH2, SFRP1, and WNT5A. (A) The miR26a level in C4-2B and C4-2B EnzR cells. n = 3, mean ± SD, ***p < 0.001, one-way ANOVA. (B) Statistical analysis of each group. n = 5, mean ± SD, *p < 0.05, ****p < 0.0001, one-way ANOVA. (C) In vitro cell migration and invasion assays. Microscope images of anti-migration and anti-invasion effects in each group (visualized with 0.1% crystal violet, scale bar: 100 μm). C4-2B EnzR cells were incubated with miR26a mimic and miR26a inhibitor, respectively (miR26a mimic/inhibitor: 1 µg·mL^− 1). (D) Dual luciferase reporter assays were used to verify the direct targeting of miR26a on EZH2 and WNT5A, h-EZH2-3’UTR/h-WNT5A-3’UTR: 0.16 µg, hsa-miR-26a-5p/NC: 5 pmol. n = 3, mean ± SD, ****p < 0.0001, one-way ANOVA. (E) Protein expressions of EZH2, WNT5A, H3K27me3, and SFRP1 were detected via western blotting in siEZH2, siSFRP1, and their negative control groups (siNC1: EZH2 negative control, siNC2: SFRP1 negative control). (F) Statistical difference of the ratio of gray values. (G) Protein expressions of WNT5A were detected via western blotting in siEZH2 + siSFRP1 and siEZH2 + siNC1 groups. (H) Statistical difference of the ratio of gray values. n = 3, mean ± SD, **p < 0.01, ***p < 0.001, ****p < 0.0001, one-way ANOVA