Figure 8.

Binding to p35 and/or DCX is required for nestin's effect on GC size. A, Representative images of DIV2 cortical neurons expressing nestin or T316-nestin mutants compared with EV control or INA-myc. β3 tubulin and phalloidin were used to visualize MTs and F-actin, respectively. Arrows indicate axonal GCs. Arrowheads indicate secondary neurites (dendrites). Brightness was adjusted for each image for easier viewing. Insets, Transfected neurons, identified by GFP expression. B, B′, B′′, Quantification of neuronal morphology in neurons expressing different nestin constructs (as shown in A). N = 72-61 cells counted per condition in three different independent experiments. Data were analyzed with nonparametric Kruskal-Wallis one-way ANOVA test with Dunn's correction. Pairwise p values are shown for comparisons to myc EV (control). WT nestin and T316A-nestin decrease GC area and filopodia number, but T316D-nestin does not. p values of <0.05 are indicated in bold.