Figure 2.

Sestrin RNAi in adult muscles abrogates exercise-dependent increases in climbing speed and lysosomal activity. (A) RNAi against Sestrin in adult muscles (MHC GS > dSesn^RNA-A (ON) UN) reduces climbing speed (2-way ANOVA, p ≥ 0.0489), and exercise further reduces climbing speed, particularly at later timepoints (2-way ANOVA, p ≤ 0.0184 after adult day 22). Control flies respond to exercise as normal. (B) A separate RNAi construct against Sestrin in adult muscles (MHC GS > dSesn^RNA-B (ON) UN) also reduces climbing speed (2-way ANOVA, genotype effect, p ≤ 0.0046 after adult day 8). Control flies respond to exercise as normal. Climbing experiments performed in duplicate, n ≥ 100 flies. (C) RNAi against dSesn in muscle reduces exercise-induced increase in lysosomal activity in fat body (ANOVA, p = 0.0014), but MHC GS > dSesn^RNA-A (ON) flies still increase fat body lysosomal activity compared to unexercised siblings (ANOVA, p = 0.0030). (D) MHC GS > dSesn^RNA-B (ON) flies also increase fat body Lysotracker staining after exercise (ANOVA, p = 0.0372). EX = cohort after 3 weeks of ramped, daily endurance training. UN = control unexercised flies exposed to identical training environment without running. ns or lack of brackets indicates no significant difference.