Fig. 5. Effect of Notch activation on astrocyte differentiation in Dll1^lacZ/lacZ and wild-type spheres. (B) Two-day-old Dll1^lacZ/lacZ mutant (I, II, III) or wild-type (IV, V, VI) spheres were exposed to J1^EC for various time intervals as described in A. (B) Triple immunostaining involved anti-MAP2 (red), anti-GFAP (green) and TO-PRO (blue). Colocalized markers are rarely observed, and are likely to result from overlapping cells as they appear in fields that are particularly dense, and probably biologically not significant. Therefore, we consider the yellow signals (V, VI, arrows) as the presence of neurons (red) overlapping astrocytes (green). (C) Data are representative of two independent experiments. Scale bar: 50 µm.