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Fig. S1 (A) Survival curves of female daGS/+ flies +RU486 and −RU486. Experiments were started with 100 flies per condition. No significant difference in survival was detected between treatments by Log-rank test. (B) Average number of eggs laid per female after 7 days of RU486 treatment (+RU486) or untreated (−RU486). Data are presented as the mean number of eggs laid per female over a 24-h period ±SEM. Eggs were counted from ten separate vials and each vial contained ten females. No significant difference was detected between treatments by ANOVA. (C) Survival of daGS/+ flies in the presence of 20 mm paraquat after 7 days of RU486 treatment (+RU486) or untreated (−RU486). Experiments were started with 100 flies per treatment. No significant difference was detected between treatments by Log-rank test. (D) Survival of daGS/+ flies in the presence of DDT after 7 days of RU486 treatment (+RU486) or untreated (−RU486). Experiments were started with 100 flies per treatment. No significant difference was detected between treatments by Log-rank test.

Fig. S2 (A) Egg to adult development time in males and females of the indicated genotypes. Only the eclosion period of the adult flies is shown. Data are shown as percentage of flies eclosing. The delay in development observed in dfoxoΔ94 homozygotes is also observed in transheterozygotes of dfoxoΔ94/dfoxo25 and hemizygous dfoxoΔ94/Df(3R)ED5624. (Males: wDahomey, n = 114; dfoxoΔ94/+, n = 94; dfoxoΔ94/dfoxoΔ94, = 106; dfoxo25/+, n = 84; dfoxoΔ94/dfoxo25, n = 112; Df(3R)ED5624/+, n = 98; dfoxoΔ94/Df(3R)ED5624, n = 84. Females: wDahomey, n = 107; dfoxoΔ94/+, n = 106; dfoxoΔ94/dfoxoΔ94, = 116; dfoxo25/+, n = 114; dfoxoΔ94/dfoxo25, n = 118; Df(3R)ED5624/+, n = 93; dfoxoΔ94/Df(3R)ED5624, n = 99). (B) Body weights and wing areas as indicators of adult fly body size in females of the indicated genotypes. Data are presented as means ± SEM (= 10 for each measurement). The reduced body size of dfoxoΔ94 homozygotes is also observed in transheterozygotes of dfoxo25/dfoxoΔ94 and hemizygous Df(3R)ED5624/dfoxoΔ94 (**P<0.05, t-test).

Fig. S3 Effects of rapamycin treatment on the survivorship of dfoxoΔ94 mutants.

Fig. S4 Lifespan and fecundity responses of dfoxoΔ94 mutant females to dietary restriction.

Table S1 Median lifespans and statistical analysis of dfoxoΔ94 mutants and control flies across a food dilution series.

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