Fig. 3: Usp10/USP10 and rin/G3BP1 regulate mitochondrial activities.

a Standardized wing posture assay showing the effects of Usp10 and rin in the fly indirect flight muscle. 25 flies were examined per group; four groups (n = 4) were counted per genotype. Control (w-) serves as a negative control; Pink1-/Y serves as a positive control. b Standardized ATP measurements (normalized to the Control) on fly muscle samples showing the effects of Usp10 and rin on energy production. 5 male flies were used per sample; six samples (n = 6) were measured per genotype. c Transmission electron microscopy (TEM, left) and Toluidine blue staining (right) images showing the effects of Usp10 and rin on mitochondrial ultrastructure in Drosophila indirect flight muscle tissue. The boxed regions are magnified in the right panel. Five samples (n = 5) were analyzed. d BN-PAGE analysis showing the regulation of mitochondrial respiratory chain complex assembly by Usp10 and rin. Control (w-) serves as a negative control; mt:COII is a loading control for BN-PAGE. The signal intensity of the corresponding band indicates the assembly of the complex. Three experiments (n = 3) were conducted. e Mito-roGFP2 imaging in fly muscle tissue showing the mitochondrial redox status modulation by Usp10 and rin. f Quantification of images shown in (e). An increase in the 405 nm/488 nm ratio indicates elevated oxidative stress. Ten samples (n = 10) were analyzed. g JC-10 staining in fly muscle tissue showing the mitochondrial membrane potential modulation by Usp10 and rin. h Quantification of images shown in (g). An increased red /green ratio indicates elevated mitochondrial membrane potential; vice versa. Eleven samples (n = 11) were analyzed. i Immunoblotting showing changes in mitochondrial, fission/fusion, and mitophagy markers affected by Usp10 and rin. Control (w-) serves as a negative control; Actin is used as a loading control in blots. All scale bars, 5 µm, except in c One-way ANOVA test (two-sided) followed by post hoc Dunnett’s multiple comparisons test 95% confidence interval (CI) was used in (a, b, f, h). Data are means ± SEM. The p values are included in the figure. Source data are provided as a Source Data file.