Effect of Vangl2 on dendrite branching. (A) Cultured hippocampal neurons were transfected with various HA-tagged constructs of Vangl2 and EGFP at 4 DIV. Neurons were fixed and immunostained using anti-GFP and -HA antibodies at 10 DIV. (B) Neurons transfected with Vangl2 constructs were visualized by immunohistochemistry to GFP and the morphology of single neuron was extracted on ImageJ for subsequent Sholl analysis. (C, D) The dendritic complexity of neurons expressing the indicated constructs was measured by Sholl analysis. Total intersections and that at various radial distances (10-100 μm) from the center of the cell soma were significantly suppressed by the Vangl2-ΔN construct. On the other hand, neurons expressing Vangl2-ΔC showed increased dendritic complexity. Data are means ± SEM (n = 20). *p <0.05 and **p <0.01 as assessed by one-way ANOVA.