Fig. 3. Neurospheres differentiate in response to Notch signaling with different sensitivity for neurons and astrocytes. (A) Partial inactivation of Notch signaling in Dll1^lacZ/+ heterozygous spheres induces a moderate increase in neurons (MAP2, in red), a decrease in astrocytes (GFAP, in green) and dramatically alters their morphology (after 3 days of differentiation). (B) Quantitative estimation for A. Data are representative of three independent experiments. (C-E) Differentiation of Dll1^lacZ/lacZ homozygous spheres in response to various concentrations of J1^EC. (C) Experimental protocol: Dll1^lacZ/lacZ mutant spheres were allowed to differentiate in the presence of various concentrations of J1^EC. (D) Triple immunostaining using anti-MAP2 (red), anti-GFAP (green) and TO-PRO (blue). (E) Quantitative results of D. Data are representative of three independent experiments. Scale bar: 50 µm. I,II,III and IV indicate the dilutions of J1^EC.