Fig. 6

Effects of Wnt3a, Wnt5a and Wnt7a on the generation of oligodendrocytes from NSCs. (a) Scheme of the in vitro assays performed to evaluate the effects of Wnt3a, Wnt5a and Wnt7a on the proliferation and differentiation of NSCs. (b) b-Catenin labelling following the addition of Wnt proteins. (c, d) BrdU labelling and the proportion of proliferative NSCs following the addition of Wnt proteins (200 ng/mL). (e) Triple immunocytochemistry performed after 2 days of NSCs differentiation to identify neurons (b-III Tubulin, red), astrocytes (GFAP, green) and oligodendrocytes (O4, yellow), and analyzed in (f). (g) O4 labelling performed after 5 div in the presence of Wnt proteins (200 ng/mL), and analyzed in (h) as the total area occupied by O4 and, the proportion of OPCs, Immature and Mature oligodendrocytes. N = 3 independent experiments. Statistics *p ≤ 0.05; **p ≤ 0.01; ***p ≤ 0.001 vs Basal; ANOVA followed by Bonferroni´s post-hoc test. Scale bar: 25 mm in a; 50 mm in c, e, g