Figure 8.
Loss of dUsp45 extends lifespan and mitigates polyQ-induced toxicity in Drosophila. (A) Western blot analysis of cathepsin-L (CTHL) levels in the heads of adult flies at different ages. (B) Quantification of CTHL levels normalized to Tubulin in A. Data are shown as mean ± SEM of three independent experiments. (C) RT-PCR analysis of dUsp45 mRNA levels in the heads of adult flies at different ages. (D) Quantification of dUsp45 mRNA levels normalized to Actin. Data are shown as mean ± SEM of three independent experiments. (E) RT-PCR analysis of dUsp45 mRNA levels in control (w1118 and dUsp45EY/+) and dUsp45 mutant (dUsp45EY/EY and dUsp45EY/B) flies. Numbers below lanes indicate the relative ratio of dUsp45/Actin. (F) Loss of dUsp45 (dUsp45EY/EY and dUsp45EY/B) extends lifespan compared with controls (w1118 and dUsp45EY/+). N ≥ 582 flies of each genotype. (G) Confocal images of phalloidin stained retinae from young (1 day) and aged (70 days) adult flies expressing GFP (control), or mutant (72Q) Htt fragment together with GFP or dUsp45RNAi under control of the eye-specific driver GMR-Gal4. Flies were reared in a light/dark (LD 12L:12D) cycle incubator. Scale bar, 10 μm. (H) Quantification of intact rhabdomere numbers in flies described in G. Data shown as mean ± SEM in n = 3, ≥30 ommatidea per genotype. (I) Confocal images of phalloidin stained retinae from young (1 day) and aged (40 days) adult flies expressing GFP (control), or mutant (72Q) Htt fragment together with GFP or CoroRNAi under control of the eye-specific driver GMR-Gal4. Flies were reared in a light/dark (LD 12L:12D) cycle incubator. Scale bar, 10 μm. (J) Quantification of intact rhabdomere numbers in flies described in I. Data shown as mean ± SEM in n = 3, ≥20 ommatidea per genotype. (K) A schematic diagram illustrating the role of USP45 in regulating autophagy, lysosome function, and V-ATPase lysosomal localization through the modulation of actin structures via Coro1B. Significance was determined using one-way ANOVA and Tukey’s multiple comparisons test (B, D, H, and J), and log-rank test (F); *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001; NS not significant. Source data are available for this figure: SourceData F8.

Loss of dUsp45 extends lifespan and mitigates polyQ-induced toxicity in Drosophila. (A) Western blot analysis of cathepsin-L (CTHL) levels in the heads of adult flies at different ages. (B) Quantification of CTHL levels normalized to Tubulin in A. Data are shown as mean ± SEM of three independent experiments. (C) RT-PCR analysis of dUsp45 mRNA levels in the heads of adult flies at different ages. (D) Quantification of dUsp45 mRNA levels normalized to Actin. Data are shown as mean ± SEM of three independent experiments. (E) RT-PCR analysis of dUsp45 mRNA levels in control (w1118 and dUsp45EY/+) and dUsp45 mutant (dUsp45EY/EY and dUsp45EY/B) flies. Numbers below lanes indicate the relative ratio of dUsp45/Actin. (F) Loss of dUsp45 (dUsp45EY/EY and dUsp45EY/B) extends lifespan compared with controls (w1118 and dUsp45EY/+). N ≥ 582 flies of each genotype. (G) Confocal images of phalloidin stained retinae from young (1 day) and aged (70 days) adult flies expressing GFP (control), or mutant (72Q) Htt fragment together with GFP or dUsp45RNAi under control of the eye-specific driver GMR-Gal4. Flies were reared in a light/dark (LD 12L:12D) cycle incubator. Scale bar, 10 μm. (H) Quantification of intact rhabdomere numbers in flies described in G. Data shown as mean ± SEM in n = 3, ≥30 ommatidea per genotype. (I) Confocal images of phalloidin stained retinae from young (1 day) and aged (40 days) adult flies expressing GFP (control), or mutant (72Q) Htt fragment together with GFP or CoroRNAi under control of the eye-specific driver GMR-Gal4. Flies were reared in a light/dark (LD 12L:12D) cycle incubator. Scale bar, 10 μm. (J) Quantification of intact rhabdomere numbers in flies described in I. Data shown as mean ± SEM in n = 3, ≥20 ommatidea per genotype. (K) A schematic diagram illustrating the role of USP45 in regulating autophagy, lysosome function, and V-ATPase lysosomal localization through the modulation of actin structures via Coro1B. Significance was determined using one-way ANOVA and Tukey’s multiple comparisons test (B, D, H, and J), and log-rank test (F); *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001; NS not significant. Source data are available for this figure: SourceData F8.

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