Fig. 3

Drp1 inhibition attenuates autophagic blockade and protein aggregation in dopaminergic N27 neuronal cells with inducible α-syn. a & b Stable N27 cells were transfected with siRNA-Drp1 or scramble control for 24 h, then induced with PonA for an additional 48 h, fixed and immediately incubated with Proteinase-K (PK) for digestion. Drp1 and α-syn were then immunostained (arrows pointing to the characteristic PK resistant α-syn puncta) c PK resistant α-syn punta were quantified using Image J. d & e Stable N27cells were co-transfected with LC3-cherry plus either siRNA-Drp1 or scramble control for 24 h, then induced with PonA for an additional 48 h. Cells were then immunostained for α-syn and p62. f LC3 and p62 punta were quantified using Image J. Data represents mean ± SEM, n = 3–4 independent experiments with 20–30 cells per treatment group were counted in each experiment, analyzed by one-way ANOVA, followed by Newman-Keuls post hoc test. *p < 0.05. *p < 0.05 Scale bar: 10 μm