Mitochondrial Morphology in Drosophila Muscle

For the imaging experiments, we did not observe any sex-based difference in measurements, therefore we used both male and female Mef2-Gal4 > UAS-mitoGFP flies, which is aligned with a previous study that used the same line^{34 (link)}. Mef2-Gal4 > UAS-mitoGFP flies (both male and females) were anaesthetized by CO2, and their adult thorax muscles, which are commonly used in examining mitochondrial morphology and respiration^{32 (link),34 (link)}, were dissected and stained as described in Hunt and Demontis^{68 (link)}. We used rabbit anti-Mef2 (1:200, Eileen Furlong, Heidelberg, Allemagne), Alexa-conjugated phalloidin (1:200, Thermo Fisher, Waltham/Massachusetts) and goat anti-GFP (1:200, Abcam, ab6673). Samples were acquired on Leica SP5 microscopes (Biosit, IGDR-Rennes) at 40X magnification and 1024/1024-pixel resolution. Images were processed with ImageJ software.