Fig. 1

α-Synuclein expression is associated with increased levels of Ub^G76V-GFP in HEK cells and in primary cortical neurons. a Schematic of the UPS reporter. Expression of Ub^G76V-conjugated GFP is driven by the CAG promoter, a hybrid of the CBA and CMV promoters. b Representative western blot demonstrating Ub^G76V-GFP accumulation in response to MG132 treatment and A53T α-synuclein over-expression in HEK293 cells. c Primary rat cortical neurons were infected with AAV-Ub^G76V-GFP and either AAV-Empty or AAV-A53T at 3000 MOI at day 2 in vitro and fixed 6 days later. Representative images of anti-GFP (green) and anti-α-synuclein (red) immunofluorescent staining with DAPI nuclear stain (blue) are shown. Scale bar 5 μm. d Cultures infected with AAV-A53T α-synuclein display a 31% increase in mean neuronal GFP staining intensity relative to those infected with AAV-Empty vector control (**p < 0.01; n = 3 independent experiments, analyzing a minimum of 10 cells per condition)