Fig. 7: Aspirin-induced α-syn clearance is dependent on K63 ubiquitination in PD mice.

a Experimental scheme for aspirin treatment in a PD mouse model overexpressing different ubiquitin variants. AAV vectors expressing three ubiquitin variants (WT, K48R, K63R) were introduced to PD mice. 6 mice were used for each of the PFF-WT-Ub, the PFF-K48R-Ub, and the PFF-K63R-Ub treatments, respectively. The figure was created using BioRender.com. Open field test (b), pole test (c) and rotarod test (d) for PD mice with different ubiquitin variants. Each dot represents a mouse (n = 6, One-way ANOVA with FDR correction for multiple comparisons, data presented as mean ± SEM). e The K63R ubiquitin significantly weakens the clearance effect of p-α-syn caused by aspirin in the striatum. Representative immunofluorescence images of the striatum (scale bar, 100 μm) are shown. Zoom-in merged images are shown on the right (scale bar, 10 μm). Quantitative results are shown in f–h, and the dot represents the mouse brain. Impact of different ubiquitin variants, when treated with aspirin, on p-α-syn intensity (f) and density (g), as well as DAT intensity (h) in the striatum (n = 5 or 6, One way ANOVA with FDR correction for multiple comparisons, data expressed as mean ± SEM). i The K63R ubiquitin significantly decreases the number of DA neurons in the substantia nigra. Shown are representative immunofluorescence images of the substantia nigra (scale bar, 200 μm). Zoom-in of the merged images are shown on the right (scale bar, 20 μm). Quantitative results are shown in j–m, and the dot represents the mouse brain. Impact of different ubiquitin variants, when treated with aspirin, on p-α-syn intensity (j) and density (k), as well as DAT intensity (l) and DA neuron number (m) in the substantia nigra. The number of DA neurons was determined by counting DAT-positive cells in the substantia nigra (n = 4, 5 or 6, One way ANOVA with FDR correction for multiple comparisons, data expressed as mean ± SEM). Source data are provided as a Source Data file.