Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2021 Sep 19;10(9):2479. doi: 10.3390/cells10092479

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© 2021 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

PMC Copyright notice

AKT is required for Sestrin and exercise to improve climbing speed and enhance lysosomal activity. (A) Flies overexpressing dSesn in muscles (Attp;Mef2 > dSesn^WT) climb faster than controls and climb as fast as exercised controls (2-way ANOVA, p ≤ 0.0156, p ≥ 0.1300). Exercised flies expressing RNAi against AKT in muscles (Mef2 > Akt^RNAi UN) improve climbing speed but do not reach the speed of exercised controls (2-way ANOVA, p ≤ 0.0319). dSesn expression does not rescue climbing speed (Mef2 > dSesn^WT; Mef2 > Akt^RNAi) (2-way ANOVA, genotype effect, Mef2 > Akt^RNAi UN vs. Mef2 > dSesn^WT; Mef2 > Akt^RNAi UN, p ≤ 0.0463). (B) Flies expressing RNAi against AMPKα in muscles (Mef2 > SNF1A^RNAi) have low climbing speed and do not improve with exercise (2-way ANOVA, genotype effect, p ≤ 0.0070 after day 12, exercise effect, p ≥ 0.4340). dSesn expression does not rescue climbing speed to the level of Attp;Mef2 > dSesn^WT flies in flies with muscle-specific AMPKα RNAi (Mef2 > dSesn^WT;Mef2 > SNF1A^RNAi) (2-way ANOVA p ≤ 0.0269 after day 12). Climbing speed experiments performed in duplicate or triplicate, n ≥ 100 flies. (C) Fat body Lysotracker staining is low in both Mef2 > Akt^RNAi and Mef2 > dSesn^WT; Mef2 > Akt^RNAi flies whether exercised or not (compare to Attp;Akt^RNAi EX, ANOVA, p < 0.0001). (D) Exercised Attp;SNF1A^RNAi, Mef2 > SNF1A^RNAi, and Mef2 > dSesn^WT;Mef2 > SNF1A^RNAi flies increase fat body Lysotracker staining with exercise (ANOVA, p < 0.0001). (E,F) Myocardial Lysotracker staining is similar in background controls and Mef2 > Akt^RNAi and Mef2 > SNF1A^RNAi flies independent of both wild-type dSesn overexpression and exercise. Representative 63× images of Lysotracker staining in fat body (left panels) and myocardium (right panels) of exercised and control flies expressing muscle-specific RNAi against (G) Akt and (H) AMPKα, with and without muscle-specific dSesn expression. 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 represent significant 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.