Figure 10.

The γC3 variable cytoplasmic domain is necessary and sufficient to rescue dendritic arborization in C3KO neurons. A, Schematic depiction of γC3 deletion transfection constructs transfected. Asterisk indicates palmitoylation sequence at the N terminus of the γC3-VCD construct. B, Quantification of area under the curve for Sholl analysis of C3KO cultured cortical neurons transfected with GFP (control), a full-length (FL) γC3 construct, constructs lacking either the extracellular domain (C3Δecto) or the constant domain (C3Δcon), or a construct encoding only a palmitoylated γC3 variable cytoplasmic domain (C3-VCD) and analyzed at DIV8. All constructs significantly rescued arborization toward wild-type levels (indicated by the dashed line), indicating that the C3 VCD is necessary (Fig. 9) and sufficient. n ≥ 25 neurons per condition. One-way ANOVA; Dunnett's multiple comparisons test. Error bars represent the SEM; ****p < 0.0001. C, Representative traces of neurons from each transfection condition. D, Representative images of transfected neurons immunostained for GFP (transfected to fill neuronal processes) and the HA tag on the indicated construct. All constructs localize in a punctate or patchy manner in neuronal dendrites. Scale bar: 50 µm (C) or 10 µm (D).