Figure 2.

PrP^Sc in the purified fractions from scrapie and GSS-A117V. P2 pellets obtained from scrapie affected voles (Bv-Scr) (A) or GSS-A117V (B) with the purification methods using PE (PE_P2) or PK (PK_P2) as proteases, were analysed by western blot either untreated (–) or subjected to a further PK digestion (+). The PE_P2 fraction from Bv-Scr contained full-length PrP^Sc that upon PK digestion showed the typical three band PrP^res pattern. In contrast, the GSS-A117V PE_P2 fraction contained low molecular weight C-terminal cleaved PrP^Sc fragments, not detected by the C-terminal antibody SAF60, but partially preserving the octarepeat region in the N-terminus (detected with SAF32). Upon PK digestion of PE_P2 from GSS-A117V, PrP^res was only detected with antibody 9A2, revealing the expected 7 kDa PrP^res profile, devoid of both the N- and C-terminus. The GSS-A117V PK_P2 pellet contained only 7 kDa PrP^res, already devoid of N-terminal and C-terminal moieties, as it was only detected by the internal antibody 9A2.