Figure 2.

H3.3 regulates the proliferation of the NSCs. (a,b) H3.3 knockdown causes GFP-positive cell positioning changes in embryonic brain. The empty shRNA control or H3.3KD plasmid was electroporated into E13.5 embryonic mouse brains, and the phenotype was analyzed at E17.5. The percentage of GFP-positive cells in each region is shown (n=3 independent experiments; error bars represent mean±S.E.M; *P<0.05). The scale bar represents 50 μm. (c,d) BrdU incorporation is reduced in H3.3 knockdown plasmids-electroporated sections. The mouse brain was electroporated at E13.5 and BrdU (50 mg/kg) was injected 2 h prior to killing at E17.5. The white arrows indicate GFP and BrdU double-positive cells. The bar graph shows the percentage of BrdU and GFP double-positive cells relative to the total GFP-positive cells in the VZ/SVZ (n=3 independent experiments; *P<0.05, bars represent mean±S.E.M). The scale bar represents 50 μm. (e,f) The percentage of GFP+BrdU+PAX6+ cells is decreased in H3.3 knockdown brains. The mouse brain was electroporated at E13.5 and BrdU (50 mg/kg) was injected 2 h prior to killing at E17.5. The white arrows indicate GFP+BrdU+PAX6+ cells. The bar graph shows the percentage of GFP+BrdU+PAX6+ cells relative to the total GFP-positive cells in the VZ/SVZ (n=3 independent experiments; *P<0.05; bars represent mean±S.E.M). The scale bar represents 20 μm