Fly lifespan analysis.

For survival analysis, flies were collected within 24 hours from eclosion, sorted by sex under light CO₂ anesthesia, and reared at standard density (20–25 flies per vial) on cornmeal, soy flour, and yeast fly food at 25°C and 60% humidity and a 12-hour light/12-hour dark cycle. Flies were transferred to fresh vials every 2 days and dead flies counted. RU486 (Mifepristone) dissolved in ethanol was administered in the media at a final concentration of 150 μg/ml. The efficiency of Smn and p62 RNAi lines was tested using the ubiquitous actin5c-Gal4 driver. As a control, we used RNAi against the white gene, responsible for the eye color that we and others have showed does not affect lifespan when it is mutated or suppressed with RNAi (79). The following RNAi lines were used: control RNAi (HMS00017), Smn-i (JF02057), p62-551i (HMS00551), and p62-938i (HMS00938).