Fig. 1. Loss of UCHL1 rescues the PD-related phenotypes of PINK1 and Parkin mutants in Drosophila.

(A) Genetic scheme to screen new suppressive genes for the swollen mitochondrial phenotypes of PINK1 (PINK1^B9) and Parkin (park¹) null mutants. (B) Confocal fluorescence images of the adult flight muscles. Green indicates mitochondria, and red indicates actin filament. Scale bar, 5 μm. Percentage of the flies having swollen mitochondria was quantified in fig. S3C. (C) Measurement of the climbing ability in the adult flies. n = 10. (D) Confocal immunofluorescence images and numbers of DA neurons in the PPM1/2 regions of adult fly brains. n = 10. Green indicates DA neuron. Scale bar, 20 μm. The number in panels indicates the number of DA neurons in each image. (E) Measurement of the DUB activities of UCH mutant proteins. Linear graphs show relative DUB activities of the mutants normalized by the activity of UCH WT at 30 min. Bar graphs show area under the curve (AUC) of the linear graphs. n = 3. (F) Confocal fluorescence images of the adult flight muscles. Green denotes mitochondria, and red denotes actin filament. Scale bar, 5 μm. Percentage of the flies having swollen mitochondria was quantified in fig. S5A. (G) Confocal fluorescence images for terminal deoxynucleotidyl transferase–mediated deoxyuridine triphosphate nick end labeling (TUNEL) assays of the adult flight muscles. Green denotes apoptosis, and blue denotes nucleus. Scale bar, 5 μm. Percentage of the flies showing apoptotic responses was quantified in fig. S5B. (H) Confocal immunofluorescence images (H) and numbers (H′) of DA neurons in the PPM1/2 regions of adult fly brains. n = 10. Green indicates DA neuron. Scale bar, 20 μm. The number in panels indicates the number of DA neurons in each image. Two-way analysis of variance (ANOVA) with Sidak’s multiple comparison test was used (C and D), and one-way ANOVA with Dunnett’s multiple comparison test was used (H′). Data were presented as means + SD.