Figure 4.

miR‐96 and miR‐467a‐3p target the Sox6 to regulate the neural differentiation. A, The expression of Sox6 during the neural differentiation from ESCs to NPCs. B, Target validation of the binding of Sox6 3′UTR by miR‐96 or miR‐467a‐3p. C, Luciferase report assay indicated that miR‐96 or miR‐467a‐3p targeted wild‐type Sox6 3′UTR but not mutant UTR. D, Over‐expression of miR‐96 or miR‐467a‐3p decreased the protein level of Sox6. E, The Sox6 restored the neural differentiation repression and the Sox1‐positive cells decrease caused by over‐expression of miR‐96. F, The quantification of Sox1‐positive cells using FACS indicated that over‐expression of miR‐96 reduced Sox1‐positive cells, and that Sox6 mitigated such reductions. G, RT‐PCR showed that the mRNA levels of Sox1 and Nestin repressed by over‐expression of miR‐96 were also rescued by Sox6. H, The Sox6 restored the neural differentiation repression and the Sox1‐positive cells decrease caused by over‐expression of miR‐467a‐3p. I, The quantification of Sox1‐positive cells using FACS indicated that over‐expression of miR‐467a‐3p reduced Sox1‐positive cells, and that Sox6 mitigated such reductions. J, RT‐PCR showed that the mRNA levels of Sox1 and Nestin repressed by over‐expression of miR‐467a‐3p were also rescued by Sox6. The scale bar represents 100 μm. Ctrl, control; Data are represented as mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001; by one‐way ANOVA (A, C, F, G, I and J). n = 3 independent experiments