We purchased all drugs from Sigma-Aldrich. In the pirenperone experiment, drug was initially dissolved in 100% dimethyl sulfoxide (DMSO) at 10 mM concentration, aliquoted and stored at −20°C. Prior to the experiment, 200 µM pirenperone and the same dilution of DMSO (control vehicle) were made. Then, 50 µl of the diluted drug or control vehicle was added to each food well. After the liquid evaporated (~2 hr), the food plates were ready for use as described in Survival assays.

In the GeneSwitch experiment, RU486 (mifepristone) was first dissolved in 80% (v/v) ethanol at 10 mM concentration, marked by blue dye (5% [w/v] FD and C Blue No. 1; Spectrum Chemical), and stored at −20°C. For experimental food, 200 µM RU486 or the same dilution of control vehicle (80% [v/v] ethanol) was made from the stock and added to the food. To ensure flies consumed the same amount of RU486 between the choice diet and the fixed diet, drug was only administrated in the yeast portion of the choice diet, since the yeast intake on a choice diet is indistinguishable from the sugar–yeast intake on the fixed diet (Figure 1E). Lifespan assay was carried out as described below.

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