Fig. 7.

Knocking down miR‐637 reversed anti‐stemness phenotypes conferred by shcircEPHB4. (A) The expression of miR‐637 was measured by qRT‐PCR in indicated LN229 cells. (B) The viability of indicated LN229 cells was measured by MTT assay for 1, 2, 3, 4 and 5 days, respectively. (C,D) The stemness of indicated cells was examined by neurosphere formation assay, with representative images of formed neurospheres shown in (C) and the quantification results in (D). (E,F) The expression of CD133 on the surface of indicated cells was examined by flow cytometry in (E) and the quantification results are shown in (F). (G,H) The expression levels of the stem‐cell markers Nestin, Oct4, Nanog, CD133 and CD44 were measured by Western blot in indicated cells (G) and quantified as the ratio to GAPDH (H). LN229 cells were simultaneously manipulated for the levels of circEPHB4 and miR‐637 (shcircEPHB4 + miR‐637 inhibitor). Data are presented as mean ± SD from three independent experiments. Comparison between three or more groups was performed using one‐way ANOVA followed by Tukey’s post hoc test. *P < 0.05, **P < 0.01 and ***P < 0.001.