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. 2020 Apr 17;143(5):1512–1524. doi: 10.1093/brain/awaa078

Figure 1.

Figure 1

PrP species in PK-treated and untreated inocula. Total PrP (first lane of each blot) and PrPres (second lane of each blot) in brain homogenates from GSS-A117V, sCJD-MM1 and Bv-CWD were analysed by western blot using monoclonal antibodies directed to the central region (9A2) or the C-terminus (SAF60 and SAF84 for human and bank vole PrP, respectively) of PrP. The second lane of each blot was duplicated as a schematic and added on the right of the blot to better illustrate the PrPres electrophoretic pattern of the PK-treated samples analysed; the size of each rectangle represents the amount of chemiluminescent signal of the respective PrPres fragment in the blot. The PK-treated GSS-A117V inoculum was characterized by 7 kDa PrPres devoid the C-terminus of the protein, as it is recognized by the internal antibody 9A2 but not by the C-terminal antibody SAF60 (larger rectangle in the 9A2 schematic box). Note the presence of multimers of 7 kDa PrPres (higher molecular weight bands of ∼14 and ∼21 kDa, represented by the thinner rectangles in the 9A2 schematic box), which share the same antibody reactivity of the 7 kDa PrPres, i.e. they are recognized by the internal antibody but not by the C-terminal one. The PK-treated sCJD-MM1 inoculum was characterized by a complex PrPres pattern, resulting from the overlapping of (i) type 1 PrPres, recognized by both the antibodies (three dark grey rectangles in both the schematic boxes), which is characterized by a dominance of the mono-glycosylated fragment, with an unglycosyilated fragment of 21 kDa in size; (ii) an additional C-terminal, fully glycosylated PrPres fragment that is recognized only by the C-terminal antibody (SAF60, three light grey rectangles in the SAF60 cartoon box), with an unglycosylated fragment of 14 kDa in size. The PrPres in the Bv-CWD inoculum was instead characterized by a classical PrPres pattern with an unglycosylated fragment of 19 kDa in size and a dominance of the di-glycosylated fragment (9A2 and SAF84, three dark grey rectangles in both the schematic boxes). The asterisks indicate non-specific bands detected in untreated brain homogenates by the antibody 9A2.