Biochemical traits of PrPSc of C- and L-BSE prions accumulated in the brains.

(A) Western blot analysis. Homogenates of bovine and monkey brains were digested by PK, and subjected to SDS-PAGE using a Bis-Tris/MES-NaOH buffer system (top panel). Alternatively, PK digests were further treated by PNGase F, and subjected to SDS-PAGE using a Bis-Tris/MES-NaOH buffer system (middle) or a Tris-HCl/Tris-glycine buffer system (bottom). Monkeys #007 and #011 were infected by bovine C-BSE prions; monkeys #014 and #015 were infected by bovine L-BSE prions. Mr; Relative molecular mass. (B) Ratios of signal intensities of di-, mono-, and non-glycosylated forms of PrPSc by Western blot analysis. PrPSc was detected using a LAS-3000 mini imaging system. Data shown are from four independent analyses. (C) Western blot analysis showing PK sensitivity of PrPSc of bovine C- and L-BSE prions. Brain homogenates were incubated with PK at the indicated concentrations (37°C for 30 min). PrPSc was detected using the LAS-3000 mini imaging system, and signal intensities of PrPSc were plotted relative to those detected in samples digested with 50 μg/mL of PK, which were set at 100.0%. The data are from three independent analyses. (D) PK sensitivity of PrPSc of monkey C- and L-BSE prions, as in (C). The data are from three independent analyses of homogenates of the brains of #007 and #015. In (A), (C), and (D), the anti-PrP antibody SAF-84 was used.