Aggregation and toxicity of α-synuclein in HEK293T cells.

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Aggregation and toxicity of α-synuclein in HEK293T cells. Aggregation and toxicity of α-synuclein in HEK293T cells. A, B, Aggregation of wild-type and mutant α-synuclein in HEK293T cells transfected with equal amounts of expression vectors encoding wild-type and mutant α-synuclein. A, Two days after transfection, cells were fixed and immunostained with antibodies against the myc-epitope. B, The number of immunopositive aggregates per field was quantitated and compared with wild-type levels (means ± SEMs; ***p < 0.001 by Mann–Whitney U test; n = 3 independent cultures). C, D, Expression of wild-type and mutant α-synuclein (α-Syn) in HEK293T cells. Two days after transfection, expression levels were analyzed by immunoblotting with antibodies against the myc-epitope (C), normalized to β-actin levels, and quantitated as percentage of wild-type levels (D; means ± SEMs; n = 4 independent cultures). E, Metabolic activity of HEK293T cells transfected with wild-type and mutant α-synuclein. Two days after transfection, cells were subjected to an MTT assay. Data were normalized to wild-type α-synuclein levels (means ± SEMs; *p < 0.05 by Mann–Whitney U test; n = 6 independent cultures). Jacqueline Burré et al. J. Neurosci. 2015;35:5221-5232 ©2015 by Society for Neuroscience