Figure 8.

Raptor regulates astrocytic differentiation in vitro. A, Cells from dissociated neurospheres after 5 d of differentiation. Cells were stained with antibodies against β-tubulin III (Tuj1; red), GFAP (green), and DAPI (blue). B, Relative number of neurons (left) and astrocytes (right) derived from dissociated neurospheres. C, Western blot analysis of lysates from dissociated neurosphere cultures grown under the same conditions as in A probed with antibodies against the proteins indicated. D–F, Analysis of differentiation in E19.5 rptor^fl/+ and rptor^fl/fl neurospheres transduced with Cre-expressing adenoviruses. D, Micrographs of neurosphere-derived cells after differentiation. After 5 d, cultures were stained with Tuj1 antibody (red), anti-GFAP antibody (green), and DAPI (blue). E, Quantification of the fraction of neurons (left) and astrocytes (right) in percentage of control. F, Western blot analysis of lysates from cultures grown under the same conditions as in D probed for the indicated marker proteins. Sample number in E is 4 mice per genotype. Error bars indicate mean ± SD from three independent experiments. *p < 0.05 (two-tailed t test). **p < 0.01 (two-tailed t test). ns, Not significant (p > 0.05). Scale bars: A, D, 100 μm.