a-Syn and Innate Immunity Figure 1. Determination of the purity, oligomeric state and cytotoxic effects of preparations of a-synucleins. (A) SDS-PAGE electrophoresis and (B) native PAGE electrophoresis of wild type (Wt) and mutant (A53T, A30P and E46K) a-Syn preparations; (C) BN-PAGE of wild type (Wt) and mutant (A53T, A30P and E46K) a-Syn preparations; (D) a-Syn oligomer content after 20 hr incubation with cells (on a protomer basis), relative to the initial amount of exogenously added a-Syn, as determined by sandwich ELISA. (E) LDH release in mixed glial cultures following incubation for 20 hours with the highest concentration of a-Syn used in our experiments, i.e. 5 mg/ml. MG132, a proteasome inhibitor, was used at 6 mM as a control. Values are means from triplicate measurements. doi:10.1371/journal.pone.0013481.g001 PLoS ONE | www.plosone.org 3 October 2010 | Volume 5 | Issue 10 | e13481