TUBB3 is specifically involved in netrin-1/UNC5C-regulated MT dynamics in the GC of P2 cerebellar EGL neurons. A–E, P2 mouse EGL neurons were cotransfected EB3-GFP with control shRNA (A), UNC5C shRNA (B), UNC5C shRNA plus wild-type human UNC5C (C), TUBB3 shRNA (D), or TUBB3 shRNA plus wild-type human TUBB3 (E), respectively, and a Dunn chamber axon guidance assay was performed. A, D, E, Left, Live-cell imaging tracking EB3-GFP comet movements in the GC. Right, Kymographs of EB3-GFP comets in the proximal (upper) and distal (lower) regions of the GC. B, C, Kymographs of EB3-GFP comet movements in the proximal (upper) and distal (lower) regions of the GC of P2 cerebellar EGL cells transfected with UNC5C shRNA (B) or UNC5C shRNA plus wild-type human UNC5C (C). Scale bar, 10 μm. White triangle represents either a control or netrin-1 gradient. F, Quantification of the velocity (left) and travel distance (right) of moving EB3-GFP comets in the GC of P2 EGL cells (A–E) at 40 min after image capture. Data are mean ± SEM (3 GCs in each group). *p < 0.05 (one-way ANOVA with Tukey's test for post hoc comparisons). ***p < 0.001 (one-way ANOVA with Tukey's test for post hoc comparisons).