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

Figure 4.

Figure 4

Distinct aggregation behavior of human and mouse α-syn proteins after seeded PMCA in the different brain substrates. A–C) Immunoblots of α-syn in PMCA samples obtained after seeding the 4 mouse brain substrates with a PMCA product (dilutions 10−3 and 10−4) produced in M83+/− brain, as shown in Fig. 1A. 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 immunoblot membranes were revealed by MJFR1 or D37A6 antibodies specific for the human or mouse α-syn, respectively, or by C-20R antibody, which recognizes both human and mouse α-syn. Bars to the left of each panel indicate the ∼20-, 30-, 40-, 50-, and 60-kDa MW markers. Rec, human recombinant α-syn control. D) PMCA products were loaded on MaxiSorp plates, and captured α-syn was detected using C-20-R, MJFR1, or D37A6 antibodies. Three repetitions of the ELISA test were performed. KO, knockout; WT, wild type.