Fig. 6.

CircEPHB4 directly targeted the expression of miR‐637. (A,B) The expression of miR‐637 was measured by qRT‐PCR in LN229 or SHG‐44 cells with altered expression of circEPHB4. (C) Bioinformatic analysis revealed a potential binding site between circEPHB4 and miR‐637. (D) The luciferase activity was measured by reporter assay. Wild‐type (WT) or mutated (MUT) circEPHB4 sequence was cloned into luciferase reporter construct, co‐transfected into LN229 or SHG‐44 cells with either miR‐637 mimics or inhibitor. (E) The interaction between circEPHB4 and miR‐637 was examined by RIP assay using either anti‐Ago2 or IgG antibody. The level of circEPHB4 or miR‐637 pulled down by IgG was arbitrally defined as 1. Data are presented as mean ± SD from three independent experiments. Comparison between two groups was performed using Student’s t‐test and between three or more groups using one‐way ANOVA followed by Tukey’s post hoc test. *P < 0.05, **P < 0.01.