Figure 7.

Epistatic relationship between Trap1 and parkin. (a) Trap1 expression decreases the degree of thoracic indentations in parkin mutant flies, park²⁵. Thoracic indentations were counted up to 24 h after eclosion (n>250 for each genotype). The asterisks indicate the significance (χ², two-sided, alpha<0.05). (b) The expression of Trap1 fails to rescue the climbing defect of the park²⁵ mutants. 3-day-old flies with the indicated genotypes were tested using a standard climbing assay (mean±S.D., n>120 flies per genotype). (c) Trap1 expression increases the levels of ATP in the park²⁵ mutants. The ATP levels were measured using a bioluminescent assay. Data are shown as the means±S.D. (n=4 for each genotype). The statistical significance is indicated by the asterisks (one-way ANOVA with Dunnett's multiple comparison test relative to park²⁵). (d) Trap1 expression restores the mitochondrial complex I protein levels in the park²⁵ mutant flies. Whole-fly protein lysates were used for western blotting analysis with the indicated antibodies. (e) Trap1 expression enhances the lifespan of the park²⁵ mutants. Fly viability was scored over a period of 40 days using a minimum of 100 flies per genotype. The statistical significance is indicated by the asterisks (log-rank, Mantel–Cox test). (f) Expression of Trap1 increases the resistance of park²⁵ mutants to paraquat-induced stress. Fly viability was scored over a period of 15 days using a minimum of 80 flies per genotype. The statistical significance is indicated by the asterisks (log-rank, Mantel–Cox test). (g) The expression of parkin rescues the climbing defects of Trap1⁴ mutants. Flies with the indicated genotypes and ages were tested using a standard climbing assay (mean±S.D., n>60 flies per genotype). The asterisks indicate significant values (two-way ANOVA with the Bonferroni multiple comparison test). (h) parkin expression restores the mitochondrial complex I protein levels in Trap1⁴ mutant flies. Whole-fly protein lysates from 20-day-old flies were used for western blotting analysis with the indicated antibodies