FIGURE 3.

Aggregation kinetics of α-syn in the presence of exosomes. Aggregation kinetics for 30 μm α-syn and β-syn was followed by ThT fluorescence in the presence and absence of exosomes in 10 mm MES, pH 5.5, with 140 mm NaCl. The averages of 4–8 replicate traces are shown in boldface with individual traces dashed below. A, aggregation of α-syn (black) with 0.25 mg/ml exosomes from N2a cells (red) or N2a cells overexpressing α-syn (green) show a distinct difference in lag time. Data were collected at 100 rpm. B, aggregation of α-syn (black) with 0.25 mg/ml exosomes from cells overexpressing disease-associated mutant α-syn (purple A53T, orange A30P, and light blue E46K) also exhibit a significantly shorter lag time than α-syn alone. Data were collected at 100 rpm. C, aggregation of α-syn (black) with 0.25 mg/ml exosomes from N2a cells (red) under quiescent conditions. D, dose dependence aggregation assay of α-syn in the presence of different exosome concentrations varying from 0–0.25 mg/ml. E, lag times, corresponding to 10% of maximum intensity, taken from the conditions depicted in D pointing again toward a significantly accelerated fibrillation when in the presence of exosomes. F, control experiment with β-syn, nonaggregating α-syn homologue protein, which remained unaffected in the presence of 0.25 mg/ml exosomes during the time frame assayed.