Figure 9.

Time course of homologous and heterologous seeded fibril growth over 96 h. Homologous and heterologous mixtures of 11.5 μg/ml C2-α-syn seeds and 500 μg/ml C2-α-syn monomer were incubated at 37 °C without agitation for varying lengths of time up to 96 h. Fibril growth at each time point was determined by FlAsH fluorescence. The rate for each seed-monomer combination was compared with the rate of fibril growth for WT seeds incubated with WT monomer. A–E, time course of fibril growth for homologous seed-monomer combinations demonstrated that the rates over longer incubation periods were very similar to the rates observed in 3-h experiments (Fig. 5). Furthermore, faster and slower rates of fibril growth corresponded to higher and lower levels of fibril accumulation as rates plateaued by 96 h. F–J, time course of fibril growth for WT seeds incubated with each of the different mutant monomer species. K–O, time course of fibril growth for each of the different mutant seeds incubated with WT monomer. The time course of heterologous fibril growth over longer time periods was also similar to the rates observed in 3-h experiments (Fig. 6). P–T, time course of fibril growth when each monomer species was incubated in the absence of seeds (mean ± S.E. (error bars), n = 3, independent experiments = 2).