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. 2015 Mar 30;208(7):931–947. doi: 10.1083/jcb.201404066

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© 2015 West et al.

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Figure 9. — Synaptic overgrowth and associated up-regulated markers induced by CHMP2B^Intron5 expression are alleviated by Rab8 expression. (A and B) Panneuronal (n-Syb–Gal4) expression of the UAS-CHMP2B^Intron5 mutant transgene elicited significant synaptic overgrowth, muscle 4 hemisegment A3. Overgrowth was rescued by coexpression of UAS-Rab8. ANOVA: F(d.f. 3) = 25.2705; P < 0.001 with post-hoc Dunnett’s comparison to wild-type control: **, P < 0.01. Post-hoc Student’s t test comparison between groups: ###, P < 0.001; #, P < 0.05. (C–E) Panneuronal (n-Syb–Gal4) expression of the UAS-CHMP2B^Intron5 mutant transgene elicited a significant increase in nuclear P-MAD (C), Dad-LacZ (D), and Puc-LacZ (E) within Eve-positive motor neuron nuclei. In all cases, coexpression of UAS-Rab8 was sufficient to rescue to wild-type levels. (C) ANOVA: F (d.f. 3) = 18.873; P < 0.001. (D) ANOVA: F (d.f. 3) = 77.984; P < 0.001. (E) ANOVA: F (d.f. 3) = 93.059; P < 0.001 with post-hoc Dunnett’s comparison to wild type controls: ***, P < 0.001; **, P < 0.01. Post-hoc Student t test comparison between groups: ###, P < 0.001; ##, P < 0.01; #, P < 0.05. (F) Panneuronal (n-Syb–Gal4) expression of the UAS-CHMP2B^Intron5 mutant transgene induced accumulation of the ESCRT-III component Shrub in axons. Accumulations colocalized with both spin and Wit. Aggregates were not observed in wild-type animals. (G) Mouse primary cortical neurons transfected with CHMP2B^Intron5 show CHMP2B^Intron5 aggregates colocalizing with POSH within neuronal projections. Arrowheads denote points of colocalization between POSH and CHMP2B^Intron5. Numbers above bars = n. Error bars show SEM. Bars: (A and G) 10 µm; (F) 5 µm.