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. 2019 Aug 1;33(11):12073–12086. doi: 10.1096/fj.201900354R

Figure 5.

Figure 5

Detection of α-syn PMCA seeding activity in the brain of sick M83 mice. A) Western blot analysis of PMCA products obtained after seeding with serial 10-fold dilutions (10−3 to 10−6) of a brain homogenate from a sick M83+/+ mouse in comparison with unseeded NBH substrate treated by PMCA (NBH) or after seeding with unsonicated healthy M83+/+ brain at dilution 10−3 (CTL). In total, 144 PMCA cycles in 10% brain homogenates were performed for the C57Bl/6-KO, C57Bl/6-WT, and D substrates, or 96 PMCA cycles in 5% brain homogenates for the M83+/− substrate. The sick M83 mouse was a 170-d-old mouse that had been intracerebrally inoculated at 8 wk old with brain homogenate of sick M83 mouse, and the healthy mouse was a 60-d-old unchallenged mouse. B) Immunoblots of α-syn (C-20R antibody) in PMCA samples obtained after seeding the C57Bl/6-WT brain substrate (10%, 144 cycles) with a whole-brain homogenate (dilutions 10−3 to 10−10) from a sick M83 mouse that died with typical clinical signs of the disease in comparison with unseeded NBH. One or 2 PMCA rounds were performed. Bars to the left of each panel indicate the ∼20-, 30-, 40-, 50-, and 60-kDa MW markers. KO, knockout; Rec, human recombinant α-syn control; WT, wild type.