Figure 2

C‑BSE prions emerge during the in vitro propagation by PMCA of brain isolates from cows inoculated with atypical scrapie. A PMCA reaction products from the frontal cortex (Fc), thalamus (Th) and cerebellum (Cbl) from the four atypical scrapie-challenged cows and a negative control cow were subjected to PrP^Sc detection by western blot. Positivity was detected in Th and Cbl samples from Bovine 1; Fc and Cbl samples from Bovine 2; and Fc and Th samples from Bovine 3. No positive reactions were detected in PMCA reaction products from the negative control. PMCA reaction products from a second negative control cow were also subjected to PrP^Sc detection by western blot, showing no positive results (results not shown). A classical scrapie isolate (Scrapie + C) and a C-BSE isolate were used as positive controls for western blot. B Relative amounts of diglycosylated (black bar), monoglycosylated (grey bar) and unglycosylated (clear bar) glycoforms of PMCA products seeded with brain samples from bovines challenged with atypical scrapie and six C-BSE controls were determined by Image J software. Results are presented as mean ± standard deviation. No significant differences in the relative amount of PrP^Sc glycoforms were observed between samples from the study and C-BSE controls.