Fig. 2. Compared differentiation potential of wild-type and Dll1^lacZ/lacZ mutant neurospheres. (A) Kinetics of differentiation were analyzed for wild-type (I, III, V, VII) and Dll1^lacZ (II, IV, VI, VIII) spheres. Three-day-old spheres were fixed after 5 hours (I, II), 10 hours (III, IV), 48 hours (V, VI) and 6 days (VII, VIII) on polyornithine. Markers used for immunostaining were: anti-MAP2 (I-VI) coupled to Cy3, for neurons (red); anti-PDGFR (I-IV) coupled to Alexa 488, for OPCs (green); anti-GFAP (V, VI) coupled to Alexa 488, for astrocytes (green); and anti-O4 coupled to Cy2 (green) (VII, VIII). In all cases, nuclei were visualized by TOPRO (blue). (B) Approximate quantification of the results. Each of the cell type was quantified at various differentiation times: MAP2-positive cells after 48 hours; GFAP-positive cells after 48 hours; oligodendrocytes were estimated from PDGFR-positive cells after 10 hours on polyornithine. The data were cumulated and expressed as percentages of total cell number estimated from TO-PRO staining. Scale bar: 50 µm.