Fig. 1
A Survival of male and female flies following knockdown of Coq8 with elav-GAL4, which drives expression of Coq8 RNAi in all neurons. In all graphs, black bars indicate females and grey bars indicates males. elav/ + indicates elav-GAL4 crossed to the wild-type w1118 control. Flies were raised at 18 °C. The number of live male and female progeny were counted. Error bars indicate the standard error of three separate experiments. Female (t-test t(4) = 3.585, *p = 0.0231); Male (t-test t(4) = 16.2, ***p < 0.0001). B Flies were generated as described in A. The proportion of female flies (30/vial) to climb above 5 cm in 10 s was counted. Nine vials of flies were counted per genotype. (t-test t(16) = 23.465, ***p < 0.0001). The schematic indicates the average climbing index of the Coq8 RNAi (CI = 0.03) and control (CI = 0.7) females. C Frontal confocal projections of whole brains processed for immunohistochemistry with anti-Bruchpilot (n = 6 brains/group). Scale bar = 100 μm. D GMR-GAL4 males were crossed to UAS-Coq8 RNAi or w1118 females and the eyes of F1 progeny were analysed by light and scanning electron microscopy. Scale bar = 200 μm (top), 50 μm (bottom). E Nile red staining on dissected retinas. Scale bar 10 μm. F Dihydroethidium staining on dissected retinas. Scale bar = 10 μm. The fluorescence intensity was quantified (n = 3/group) Female (t-test t(5) = 44.68, ***p < 0.0001); Male (t-test t(5) = 3.3153, *p = 0.0211). G Flies were generated as described in E. Eyes were imaged by light microscopy at the indicated number of days post-eclosion. Scale bar = 200 μm. H The area of necrosis in each eye was quantified (n ≥ 8 eyes/group). (Female: ANOVA F(3,31) = 28.7 p < 0.0001; post-hoc Tukey’s test. Male: ANOVA F(3,32) = 27.12 p < 0.0001; post-hoc Tukey’s test, **p < 0.01, *p < 0.05). I Co-expression of mitoGFP and Myc-tagged Drosophila Coq8 with motor neuron driver D42-GAL4. Scale bar = 20 μm. J. elav-GAL4 females were crossed to males of the indicated genotypes and the number of surviving adult male F1 progeny were counted (ANOVA F(4,10) = 51.95 p < 0.0001; post-hoc Tukey’s test, **p < 0.01). K The climbing ability of surviving females was assessed (ANOVA F(4,40) = 137.46 p < 0.0001; post-hoc Tukey’s test, **p < 0.01, *p < 0.05). L GMR-GAL4 females were crossed to flies of the indicated genotypes and eyes were imaged by light microscopy 14 days post-eclosion. Co-expression of WT Coq8 rescued the necrosis resulting from Coq8 knockdown but I295P and L520* did not. M Quantification of necrosis in L (n ≥ 9 eyes/group) (Female: ANOVA F(4,43) = 7.73 p < 0.0001; post-hoc Tukey’s test. Male: ANOVA F(4,43) = 10.4 p < 0.0001; post-hoc Tukey’s test, **p < 0.01, *p < 0.05). N Co-expression of mitoGFP and human COQ8A with motor neuron driver D42-GAL4 detected with anti-COQ8A. Scale bar = 12 μm. O elav-GAL4 females were crossed to males of the indicated genotypes and the number of surviving adult male F1 progeny were counted (ANOVA F(3,8) = 133.22 p < 0.0001; post-hoc Tukey’s test, **p < 0.01). P The climbing ability of surviving females was assessed (ANOVA F(3,28) = 108.46 p < 0.0001; post-hoc Tukey’s test, **p < 0.01). Q Eyes of the indicated genotypes were imaged by light microscopy at 14 days post-eclosion. Scale bar = 200 μm. R Necrosis was quantified in the eyes of the indicated genotypes (n =  ≥ 9 eyes/group) (Female: ANOVA F(3,34) = 34.9 p < 0.0001; post-hoc Tukey’s test. Male: ANOVA F(3,34) = 77.4 p < 0.0001; post-hoc Tukey’s test, **p < 0.01)