Figure 6.

Sestrin-dependent adaptations in climbing speed and lysosomal activity depend on the TORC2 axis. (A) Flies expressing muscle specific RNAi against Sin1 (Mef2 > Sin1^RNAi) reduce climbing speed with exercise (2-way ANOVA, exercise effect, p ≥ 0.0268). Muscle-specific wild-type dSesn expression (Mef2 > dSesn^WT;Mef2 > Sin1^RNAi flies) does not rescue climbing speed. Climbing speed experiments performed in triplicate, n ≥ 100 flies. (B) Mef2 > dSesn^WT;Mef2 > Sin1^RNAi flies have low fat body Lysotracker staining whether exercised or not. (C) Myocardial Lysotracker staining is similar across genotypes independent of exercise and wild-type dSesn expression in flies with muscle specific RNAi against Sin1. (D) Representative 63× images of Lysotracker staining from fat body (upper panels, quantified in (B)) and myocardium (lower panels, quantified in (C)). Lysotracker staining n = 5, performed in duplicate or triplicate. Dotted lines in myocardial images delineate heart tissue. White triangles point to example regions of Lysotracker puncta. White asterisks indicate pericardial cells. Asterisks in histograms indicate significant pairwise post-hoc comparisons from ANOVA. Error bars depict ± SD. EX = cohort after 3 weeks of ramped, daily endurance training. UN = control unexercised flies exposed to identical training environment without running.