Figure 6. α-synuclein is essential for EV mediated neurotoxicity.
(A) EVs were isolated from the medium of cultured control HEK cells (−), HEK cells overexpressing wildtype human α-synuclein (+), and α-synuclein knockout HEK cells (KO). Proteins in the EVs and the cells that released the EVs were separated in a NuPAGE Bis-Tris gel and immunoblotted using antibodies against α-synuclein, flotillin-1, calnexin and actin. (B) Primary cortical neurons were incubated for 24 hours in the presence of EVs (20 μg protein/ml) isolated from the medium of control HEK cells (−), HEK cells overexpressing wildtype human α-synuclein (+), and α-synuclein knockout HEK cells (KO). Neuronal viability was assessed using MTS and LDH assays. Values are the mean and SEM of determinations made in 3 independent experiments. * p<0.05, ** p<0.01, ***p<0.001.