Fig. 10: Cytoskeletal toxins differentially affect dendritic arborization.

A Representative confocal images of WT or Kcnb1^R312H(+/+) neurons incubated in the absence/presence of 30 nM Cytochalasin D (CytoD). The cells were co-immunostained for actin (green) and Map2 (red) antibodies. Large clumps of actin are demonstrated in Kcnb1^R312H cells (arrows). Scale bars 20 μm. B Total dendrite length (TDL) estimated by Sholl analysis for WT (upper graph) or Kcnb1^R312H(+/+) (lower graph) neurons in the absence/presence of the indicated toxins. C Sholl intersection profiles for WT (upper graph) or Kcnb1^R312H(+/+) (lower graph) neurons in the absence (black) or presence of the indicated toxins. Within the same genotype, all toxin SIPs are statistically different from the WT SIP (P < 0.0001, Kolmogorov–Smirnov test). All measurements were taken using DIV7 neurons. Sholl analysis of individual neurons was performed using Fiji, (ImageJ). In all experiments, toxins were maintained in the media starting at DIV1 at the following concentrations: Cytochalasin D (CytoD, red), 30 nM; Jasplakinolide (Jasp, blue) 30 nM; Latrunculin A (LatA, green), 1 μM and Phalloidin (Phall, light blue), 15 nM. N = 20 neurons/genotype/experimental condition from three dams/genotype. ^*P < 0.05, ^**P < 0.01, ^***P < 0.001, and ^****P < 0.0001, one-way ANOVA with Tukey’s post hoc test.