Repo protein levels influence Drosophila lifespan and behavior. All aging in this figure was performed at 29°C. A, Lifespan of two different RNAi lines against repo and the rescue of the RNAi 1 by coexpression of a UAS-repo transgene (log–rank Mantel–Cox test p < 0.0001). B, Climbing assay of the two RNAi lines against repo and the rescue of the RNAi 1 by UAS-repo as assessed just before death (respectively 7 for RNAi 1, one-way ANOVA, Tukey's multiple-comparisons test p = 0.0001 and p < 0.0001, respectively, and 12 d for RNAi 2 at 29°C, Mann–Whitney, n = 3, p = 0.049). C, Western blot analysis of Repo levels after 7 d at 29°C. D, Sixty flies (30 females and 30 males) were used for each genotype. Only repo knock-down by the ubiquitous Actin5cGal4 or the mosaic dEAAT1-Gal4 achieved a significant detrimental effect on fly lifespan, supporting the composite nature of the effect on lifespan. The specificity of each driver is as follows: repoGal4 (pan-glial), alrmGal4 (astrocyte-like glia), NP2222Gal4 (cortex glia), moodyGal4 (subperineurial glia), MZ0709Gal4 (ensheathing glia), Actin5cGal4 (ubiquitously expressed), dEAAT1Gal4 (astrocyte-like glia, cortex glia and some subperineurial glia), and GMRGal4 (eye, used as negative control, not expressed in glial cells). ***p < 0.001.