Figure 4. Internalized αS PFF and truncated αS remain detergent-insoluble.

(A, B) Neuronally differentiated CLU198 cells were treated with 4 ug/ml of αS monomers (A) or αS PFF (B). At 0h and 24h after washing, the cell lysates were fractionated into total SDS-soluble lysates (Tot), Triton X-100 (TX-100) soluble (Sol) fraction, and TX-100 insoluble (Insol) fraction. Immunoblot analysis show that internalized αS monomers partition with the TX-100 Sol fraction (A) while bulk of αS PFF partitions with the TX-100 Insol fraction (B). (C) PCN treated with PFFs were analyzed for TotαS in TX-100 soluble (S) and insoluble (IS) fractions at 3- and 7-days post αS PFF treatment. Even at 7 days following the initial internalization of αS PFF, virtually all of the truncated αS (Δ) partitions with the insoluble fraction.