Fig. 3

The expression of miR-335-3p was activated by FoxM1. a FoxM1 mRNA level in NSCs after induced differentiation detected using RT-qPCR on days 0, 3, and 7. b FoxM1 protein level in NSCs after induced differentiation determined by Western blot analysis on days 0, 3, and 7. c FoxM1 and miR-335-3p expression in NSCs after alteration of FoxM1 examined using RT-qPCR. d FoxM1 protein level in NSCs after alteration of FoxM1 measured by Western blot analysis. e 3 sites by which FoxM1 was most likely to bind to miR-335-3p promoter region predicted by online websites. f, g Dual-luciferase reporter assay for mutant miR-335-3p promoter recombinant luciferase reporter gene vector (f) and truncated miR-335-3p promoter recombinant luciferase reporter gene vector (g) co-transfected with FoxM1 expression vector into NSCs to determine the targeting relationship of FoxM1 and miR-335-3p. h The binding relationship between FoxM1 and miR-335-3p measured using CHIP assay. The measurement data were expressed as mean ± standard deviation. The unpaired t test was employed to analyze the differences between two groups, while differences among multiple groups were analyzed using one-way ANOVA, with Tukey’s post hoc test. *p < 0.05 versus NSCs without any induced-differentiation treatment, NSCs treated with oe-NC, si-NC, or si-FoxM1 + oe-NC, or IgG antibody. The experiment was performed at least 3 times