Figure 11.

Reduced dendritic complexity of C3KO neurons can be rescued by targeted Axin1 overexpression in vitro. A, Representative images of DIV8 C3KO neurons transfected with GFP, and either an empty vector, Axin1 overexpression construct, or membrane-targeted CAAX-tagged Axin1. B, Sholl analysis graphs show dendritic crossings of spheres of increasing 10-µm intervals of neurons analyzed at DIV8. Expression of either Axin1 or CAAX-Axin1 rescues low arbor complexity proximal to the soma; only CAAX-Axin1 maintains that rescue at more distal Sholl spheres. C, Quantification of area under the curve of Sholl analysis of transfected C3KO neurons for all conditions. A significant rescue is observed with membrane-targeted CAAX-Axin1. n ≥ 25 neurons per transfection condition. One-way ANOVA; Dunnett's multiple comparisons test. Scale bar: 25 µm. Error bars represent the SEM; *p < 0.05. D, E, Western blottings of COS7 cells transfected with the indicated constructs and blotted using Axin1 and β-tubulin (loading control) antibodies. Quantification of three replicate experiments indicates no significant difference in expression levels between the two constructs.