Figure 2. Separation of PrP^C and PrP^Sc in vole brain homogenates.

A: Western blot analysis of soluble and insoluble PrP fractions from normal brain homogenate (NBH) and scrapie brain homogenate (SBH) of voles. Samples were centrifuged at 20000 g for 1 h in presence of 2% sarcosyl, and supernatants (S) and pellets (P) were analysed before (+) and after (−) PK treatment. Aliquots of samples before centrifugation (Tot) were analysed too. TE per lane were 0,2 mg for “Tot” and “S” lanes, and 0.4 mg for “P” lanes. Brackets on the left indicate the position of the FL, C1 and C2 PrP fragments. Dimers (Dim) of PrP^Sc are indicated on the right in SBH blot. B: Western blot analysis of soluble and insoluble PrP fractions from scrapie brain homogenate (SBH) and an artificially mixed sample (SBH+NBH). Samples were centrifuged as described and total (Tot), supernatant (S) and pellet (P) fractions were deglycosylated. Full-length PrP (FL), C1 and C2 PrP fragments are indicated on the left. In each lane, 0.02 mg TE were loaded. C: Western blot analysis of soluble and insoluble PrP from brain homogenates of voles infected with MM1 and MM2 sCJD. The samples were treated as in panel A and total (Tot), supernatant (S) and pellet (P) fractions were analysed with or without PK treatment. In each lane 0.3 mg TE were loaded. Brackets on the left indicate the position of the FL, C1 and C2 fragments. Dimers (Dim) of PrP^Sc are indicated on the left. A–C: Membranes were probed with SAF84. Molecular size markers are shown in kilodaltons on the right of each panel.