Expression of VPS35 pathogenic variant D620N in flies result in DA neuronal degeneration and locomotive dysfunction. (A, B) Bar graphs show number of TH-positive DA neurons in flies at 20 and 60 days after eclosion (n = 3, cohort of 10). (C) Representative confocal images of whole mount brains 60 days after eclosion of VPS35 wild type, P316S and D620N flies with magnified views of PPL1 cluster (boxed on the right). (D) Bar graph shows age-dependent climbing scores of female flies at different days after eclosion. Percentage of flies that reached the top of the column after 1 min were counted (n = 3, cohort of 20). (E) Survival curves were plotted as percentage of living flies. (n = 3, cohort of 100).