Fig. 2

GA400 is highly toxic in vivo. A Representative eye images of female flies expressing GA100, GA200, GA400 or two copies of GA200 (2xGA200 (attP40, attP2)) under the control of the eye-specific GMR-Gal4 driver. GA400 expression 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 strongly reduced eye size (One-way ANOVA + Tukey’s multiple comparisons test; n = 10 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 = 10 independent vials and 18–109 counted eggs/vial; ****P < 0.0001). D Climbing ability of adult female flies expressing GA100, GA200 or GA400 for 15 or 25 days under the control of the elav-GS driver. Climbing indices are represented as box plots and the mean is indicated by + . Circles indicate individual flies. Expression of all polyGA proteins decreased climbing ability, but induction of GA400 caused stronger changes already on day 15 (Two-Way ANOVA + Tukey’s multiple corrections test; n = 41–45 flies; age: ****P < 0.0001; genotype: ****P < 0.0001; interaction of age and genotype: P > 0.05). E Survival curves of female flies with pan-neuronal expression of GA100, GA200 and GA400 under the control of the elav-GS driver. Flies expressing GA100, GA200 and GA400 were shorter-lived than elav-GS driver control flies (****P < 0.0001; log-rank + Bonferroni’s multiple corrections test, n = 150 female flies per genotype). Flies expressing GA400 were significantly shorter-lived than flies expressing GA100 and GA200 (****P < 0.0001) and GA100 flies were significantly shorter-lived than flies expressing GA200 (****P < 0.0001). F Representative images of adult fly brains stained for anti-cleaved caspase-3 (CC3). Transgenes were induced for 30 days under the control of the elav-GS driver. Scale bar: 100 µm. G Quantification of the number of CC3-positive cells per fly brain. (One-way ANOVA + Tukey’s multiple comparisons test; n = 8–10 brains; ****P < 0.0001)