SDS-PAGE of ~14 kDa Human Recombinant Alpha Synuclein Protein Pre-formed Fibrils (Type 2) (SPR-317). Lane 1: Molecular Weight Ladder (MW). Lane 2: Alpha Synuclein Protein Pre-formed Fibrils (SPR-317).

Primary rat hippocampal neurons show lewy body inclusion formation when treated with Type 1 Alpha Synuclein Pre-formed Fibrils (SPR-322) at 4 µg/ml (D-F), but not when treated with Type 2 Alpha Synuclein Pre-formed Fibrils (SPR-317) at 4 µg/ml (A-C). Tissue: Primary hippocampal neurons. Species: Sprague-Dawley rat. Fixation: 4% formaldehyde made from PFA. Primary Antibody: Mouse anti-pSer129 Antibody at 1:1000 24 hours at 4°C; Secondary Antibody: FITC Goat Anti-Mouse (green) at 1:700 for 1 hours at RT (B, E). Counterstain: Hoechst (blue) nuclear stain at 1:4000 for 1 hour at RT (A,D). Localization: Lewy body inclusions. Magnification: 20x. C is A and B merged and F is D and E merged.

Evaluation of a-syn toxicity on primary mouse cortical neurons. Lactate dehydrogenase (LDH) is a soluble enzyme present in the cytosol that is released upon cell death. Toxicity was assessed with an LDH assay and displayed as % of vehicle control (VC). Data are presented as bar graphs and standard deviation. For statistical analysis One-way ANOVA followed by Bonferroni post-hoc test (vs VC) was used. *** p<0.001. Treatment with Type 2 PFFs did not significantly impact LDH release. Data courtesy of QPS.

Evaluation of a-syn toxicity on primary mouse cortical neurons. Mitochondrial dehydrogenase activity reduces yellow MTT to dark blue formazan crystals, a reaction catalyzed in living cells. Cell viability was assessed with an MTT assay and displayed as % of vehicle control (VC). Data are presented as bar graphs and standard deviation. For statistical analysis One-way ANOVA followed by Bonferroni post-hoc test (vs VC) was used. ** p<0.01, *** p<0.001. Treatment with Type 2 PFFs reduced cell viability (p<0.001). Data courtesy of QPS.