<i>AOX</i> does not rescue <i>pink1</i> mutants.

<p>(A) Quantitative RT-PCR using primers to <i>CG3446</i> and to <i>AOX</i>, in testes using tissue of the following genotypes <i>w</i>; <i>da-Gal4</i>/<i>+</i> (green), <i>w</i>; <i>UAS-AOX</i> (orange) and <i>w</i>; <i>UAS-AOX</i>/<i>+</i>; <i>da-Gal4</i>/+ (blue). Data normalized to housekeeping genes (see <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1002456#pgen.1002456.s006" target="_blank">Text S1</a>). (B) Quantification of fertility of <i>w pink1^RV</i> control male flies of <i>pink1^B9</i>and of <i>w pink1^B9</i>; <i>UAS-AOX</i> male flies (n = 20 animals). (C) Quantification of flight in control (<i>w pink1^RV</i>; <i>da-Gal4</i>/<i>+</i>) in <i>pink1^B9</i> mutant (<i>w pink1^B9</i>; <i>da-Gal4</i>/<i>+</i>) and in <i>pink1^B9</i> mutant flies that express <i>AOX</i> (<i>w pink1^B9</i>; <i>UAS-AOX</i>/+; <i>da-Gal4</i>/<i>+</i>). Student's t-test: ns, not significant. Error bars SEM, n = 6 experiments with 5 flies each. (D) Relative ATP levels of <i>pink1^B9</i> mutant flies (<i>w pink1^B9</i>; <i>da-Gal4/+</i>) normalized to <i>pink1^RV</i> control flies (<i>w pink1^RV</i>; <i>da-Gal4/+</i>) and of <i>pink1^B9</i> mutant flies that express AOX (<i>w pink1^B9</i>; <i>UAS-AOX/+</i>; <i>da-Gal4/+</i>) normalized to control flies that express AOX (<i>w</i>; <i>UAS-AOX/+</i>; <i>da-Gal4/+</i>). Student's t-test: ns, not significant. Error bars SEM, n = 3 experiments with 5 flies each. (E) GFP labeling of mitochondria from the adult indirect flight muscles of the following genotypes <i>w pink1^RV</i>; <i>UAS-mito:GFP</i>/<i>+</i>; <i>da-Gal4</i>/+ and <i>w pink1^B9</i>; <i>UAS-mito:GFP</i>/+; <i>da-Gal4</i>/+ and <i>w pink1^RV</i>; <i>UAS-mito:GFP</i>/<i>UAS-AOX</i>; <i>da-Gal4</i>/+. (F,G) Quantification (F) and labeling (G) of Reserve Pool (RP) vesicles in <i>pink1^RV</i> controls (<i>w pink1^RV</i>; <i>da-Gal4/+</i>) in larvae expressing the AOX transgene (<i>w</i>; <i>UAS-AOX/+</i>; <i>da-Gal4/+</i>), in <i>pink1^B9</i> mutants (<i>w pink1^B9</i>; <i>da-Gal4/+</i>) and in <i>pink1^B9</i> mutants expressing AOX (<i>w pink1^B9</i>; <i>UAS-AOX/+</i>; <i>da-Gal4/+</i>), using the stimulation protocol shown on the left. Student's t-test: * <i>p</i><0.05; ns, not significant. Error bars SEM, n>8 synapses from 4 animals. (H,I) Quantification of red to green JC-1 fluorescence (H) and examples (I) at NMJ boutons in larvae of the genotypes indicated in (F–G). Student's t-test: ns, not significant. Error bars SEM, n>16 synapses from 4 animals.