Additional file 4 of Repeat length of C9orf72-associated glycine–alanine polypeptides affects their toxicity

Additional file 4. Figure S4: Expression of polyGA from the attP40 insertion site caused comparable phenotypes to expression from the attP2 site. A-C Expression of polyGA transgenes from the attP40 insertion site under the control of the eye-specific GMR-Gal4 driver. A Representative eyes of female flies. GA400 expression, but not that of GA100 or GA200, caused a strong rough eye phenotype. B Eye size of female flies normalized to the mean of the eye size of GMR-Gal4/+ control flies. GA400 expression significantly reduced eye size (One-way ANOVA + Tukey’s multiple comparisons test; n = 7-12 fly eyes per genotype; ***P < 0.0001). C Egg-to-adult viability of flies expressing polyGA proteins under the control of the GMR-Gal4 driver. GA400 expression strongly decreased viability (One-way ANOVA + Tukey’s multiple comparisons test; n = 7-10 independent vials and 5-95 counted eggs/vial; ***P < 0.0001). D Survival curves of flies expressing single-copy transgenes of GA100, GA200 and GA400, and two copies of GA100 (2xGA100) and GA200 (2xGA200) under the control of the elav-GS driver. There was no difference in survival between flies expressing the polyGA transgenes from the attP2 or attP40 sites (P > 0.05; log-rank + Bonferroni’s multiple corrections test; n = 150 female flies per genotype). Expression of 2xGA100 or 2xGA200 reduced survival more than those of the corresponding single-copy transgenes (***P < 0.0001, log-rank + Bonferroni’s multiple corrections test; n = 150 female flies per genotype, except n = 120 for 2xGA200 flies). Expression of a single copy of GA400 reduced survival more than expression of two copies of GA100 or GA200 (***P < 0.0001, log-rank + Bonferroni’s multiple corrections test). Transgene insertion sites are indicated in brackets.