Figure 5

Tubulin polymerization is impaired in p35-expressing NPC-derived neural progeny following chemical microtubule disruption. For chemical disruption of microtubules, cultures were treated with 5 μg/mL nocodazole (Noc) for 3 hr, followed by washout and incubation in fresh differentiation media for 20 mins. NPC-derived neural progeny were fixed with glutaraldehyde and processed for β-tubulin immunofluorescence. (A,B) Chemically-induced disruption of microtubule structure in nocodazole-treated control NPC-derived neural progeny (A) was recovered by 20 mins post-washout (B). (C,D) Disruption of microtubule structure in nocodazole-treated p35-expressing NPC-derived neural progeny (C) appeared unchanged by 20 mins post-washout (D). (E) Quantitative image analysis of neurite outgrowth in NPC-derived neural progeny following treatment with nocodazole. Untreated control is shown for comparison to baseline neurite lengths. * p < 0.01 compared to vehicle-treated controls by one-way ANOVA with post-hoc Dunnett's test; # p < 0.01 compared to nocodazole-treated controls by one-way ANOVA with post-hoc Tukey-Kramer test. Scale bar = 15 μm.