Fig. 5.
E12.5 NS/P cells that lack REST do not exit the cell cycle but have significantly reduced mitotic activity. (A) Immunostaining showing that most of the NS/P cells expressing Rest shRNA express the cell division marker Ki67. GFP represents cells expressing shRNA. (B) Bar graph indicating no significant difference in the number of Ki67+ cells in NS/P cells expressing Rest or scrambled shRNA. (C) Immunostaining showing fewer NS/P cells expressing the mitotic marker phosphohistone H3 (phospho H3) when Rest shRNA is expressed. Arrows point to GFP+ cells expressing phospho H3. (D) The mitotic index (fraction of phosphoH3+/GFP+ cells) of NS/P cells expressing Rest shRNA is significantly reduced. (E) Immunostaining showing some precocious neuronal differentiation as indicated by the presence of TUJ1+ cells in NS/P cells expressing Rest shRNA (arrows). (F) Bar graph indicating low but significant number of Rest shRNA-expressing NS/P cells that are TUJ1+. Scale bars: 50 μm. All error bars are mean ± s.e.m. based on three independent experiments. *P<0.05.