Fig. 2

In vitro EC differentiation and characteristic markers: a, b Representative cell culture at day 5 and 7 respectively, labeled with anti-βIV (red) tubulin and anti-GFAP (green), in which a remarkable increase in βIV tubulin expression occurs at day 7. a′, b′. High magnification (×63) of the VZ at day 5 and 7 respectively. At day 7 the expression of βIV corresponds to multiciliated EC while at day 5 the labeling is linked to monociliated NSC. The inset in A’ shows a magnification of a monociliated NSC (arrow). The insets in B’ shows a multiciliated EC expressing GFAP (arrowhead) and a multiciliated EC not expressing GFAP (asterisk). c, c′. Representative VZ at day 7 immunolabeled with anti-S 100 β (green, associated with EC) and N-Cad (red, associated with both EC and NSC). d, d′. In vitro VZ at day 7 labeled with AQP4 (green) and β-Catenin (red). e. High magnification (×63) of the VZ labeled with EMA (mature EC marker). f. Schematic representation of the differentiation of the ependymal cells. Scale bars: a–d = 100 μm; a′, b′ = 25 μm; c′, d′ = 20 μm