Fig. 1

PFF treatment induces ps-αSyn in primary neurons. a, b Primary cortical neurons derived from wild-type (WT) and αSyn knockout (KO) mice were untreated (−) or treated with αSyn monomer (M) or PFF (F) as indicated. c, d Primary cortical neurons derived from OVX mice were untreated (−) or treated with human αSyn monomer (M) or PFF (F). e, f Primary neurons derived from wild-type mice were untreated (−) or treated with αSyn monomer (M), αSyn PFF (F), or lysozyme PFF (F/L). ps-αSyn (in a, c, and e) and total αSyn (in a and c) were detected by immunoblot analysis and GAPDH was used as a loading control. In b, d, and f, ps-αSyn was visualized by immunofluorescence staining (red), and MAP2 stain (green) was used as a neuronal marker. g Primary neurons, untreated (−) or treated with αSyn monomer (M) or PFF (F), were sequentially extracted with TBS, 1% Triton/TBS, and 2% SDS/TBS. The presence of ps-αSyn and total αSyn was detected by immunoblot analysis. h PFF-treated neurons were incubated with or without a 10-min 10 μg/mL PK digestion at 37 °C prior to immunofluorescence staining to detect ps-αSyn (red) and MAP2 (green)