Fig. 7: The mTORC2 complex interacts with the 40S ribosomal subunit recycling complex.

a Immunoblotting showing mTORC1/2 activities in Usp10 and rin OE flies. Control (w-) serves as a negative control; Actin is used as a loading control in blots. b Standardized ATP measurements (normalized to the Control) on fly muscle samples showing effects of mTOR genes in Usp10 and rin OE flies. 5 male flies were used per group; six independent groups (n = 6) were measured per genotype. c, d Immunostaining of mitochondrial markers showing the regulation of mTOR genes in Usp10 OE and rin OE flies. Control (w-) serves as a negative control. Mitochondrial morphology is visualized by mitoGFP (Usp10 group) and ATP5α (rin group). e Quantification of mitochondrial size shown in (c, d). Mitochondria counts were obtained from three samples (n = 3, 7 areas counted per sample). f PLA detection of Sin1-Flag(+)/rin(-) interaction (red puncta), showing increased binding in Usp10 OE and rin OE flies. Control (w-) serves as a negative control in (f, h, j, k). g Quantification of images shown in (f). h PLA detection of Sin1-Flag(+)/Hsc70-5(-) interaction (red puncta), showing decreased binding in Usp10 OE and rin OE flies. i Quantification of images shown in h Data were obtained from six samples (n = 6) in (g, h j). PLA detection of Sin1-Flag(+)/Marf(-) interaction (red puncta), showing increased binding in Usp10 OE flies. k Quantification of images shown in (j). l PLA detection of Sin1-Flag(+)/Drp1(-) interaction (red puncta), showing increased binding in Usp10 OE flies. m Quantification of images shown in l Data were obtained from ten samples (n = 10) in (k, m). One-way ANOVA test (two-sided) followed by post hoc Dunnett’s multiple comparisons test 95% confidence interval (CI) was used in (b, e). Two-tailed Student’s t-test was used with a 95% confidence interval (CI) in (g, i, k, m, n). A working model illustrating the interactions between the 40S ribosomal subunit recycling complex, the mTOR pathway, and ERMCS. The figure was created in BioRender¹³⁶. All scale bars, 5 µm. The p values are included in the figure. Source data are provided as a Source Data file.