Figure 3.

Gli3 effects on proliferation of neural progenitors (NPs) in the cortical midline. (A) The position that was measured for the embryonic day 15.5 (E15.5) midline cortex. (B,C) Cortical deficiency of Gli3 increased the proportion of BrdU⁺ cells vs. DAPI⁺ cells. Knockdown of miR-7 decreased the proportion of BrdU⁺ cells vs. DAPI⁺ cells. Silencing miR-7 failed to rescue the dysregulation of BrdU⁺ cells in the Gli3-deficient brain. (D,E) Cortical deficiency of Gli3 suppressed the population of cells expressing the radial glial cell (RGC) marker Pax6⁺/DAPI⁺, which was completely rescued by silencing miR-7. Silencing miR-7 showed no significant changes of the proportion of cells expressing RGC marker Pax6⁺/DAPI⁺. (F,G) Losing Gli3 increased the ratio of intermediate progenitor (IP) marker Tbr2⁺/DAPI⁺, which was restored by blocking miR-7. Knockdown of miR-7 did not affect the ratio of IP marker Tbr2⁺/DAPI⁺ at E15.5. (H,I) No difference was observed in Tbr1⁺ cells vs. DAPI⁺ cells in EG mice, which was significantly reduced by silencing miR-7. Cortical deficiency of miR-7 decreased the proportion of Tbr1⁺ cells vs. DAPI⁺ cells. The markers BrdU, Pax6, Tbr2, Tbr1 and DAPI stained for proliferative NPs, RGCs, IPs, newborn neurons and all cells, respectively. Values represent mean ± SEM. n > 9. *P < 0.05; **P < 0.01; ***P < 0.001; ns, not significant. One-way ANOVA with post hoc test was used. Scale bar = 100 μm.