Figure 4
Biochemical analysis of PrPres accumulated in the brains of ICR mice infected with TgBo-passaged Sh294. PrPres banding patterns were compared among ICR mice infected with TgBo-passaged Sh294, sheep-passaged L-BSE, mouse-adapted laboratory classical scrapie strains, and mouse-adapted C-BSE using the mAb 6H4 A and SAF84 B. Lane 1, TgBo-passaged Sh294 (first passage in ICR); lane 2, TgBo-passaged Sh294 (second passage in ICR); lane 3, TgBo-passaged Sh294 (third passage in ICR); lane 4, sheep-passaged L-BSE (first passage in ICR); lane 5, sheep-passaged L-BSE (second passage in ICR); lane 6, sheep-passaged L-BSE (third passage in ICR); lane 7, ME7; lane 8, 22L; lane 9, Chandler; lane 10, mouse-adapted C-BSE (mBSE). Laboratory strains (ME7, 22L, Chandler, and mBSE) were serially passaged in ICR mice (more than 10 passages). Lanes 1–3, 4 and 5, 6, and 7–10 in A, B were loaded with 0.5 mg, 2 mg, 0.1 mg, and 0025 mg brain equivalent, respectively. PrPres was also characterized using the mAbs SAF84 (C, lane 1–4) and 6H4 (C, lane 5–8) after PNGase deglycosylation. 1, sheep Sh294; 2, TgBoPrP mouse infected with Sh294; 3, ICR mouse infected with TgBo-passaged Sh294; 4, TgBoPrP mouse infected with L-BSE. Lanes 1–3 and 5–7 in C were loaded with 0.25 mg brain equivalent. Lanes 4 and 8 in C were loaded with 0.003 mg brain equivalent. The arrow in B shows the ~12-kDa C-terminal small fragment. Molecular markers are shown on the left of each panel. Glycoform profiles of PrPres from ICR mice infected with TgBo-passaged Sh294 (third passage), sheep-passaged L-BSE (third passage) and mBSE (14th passage) are given D. PrPres was detected with mAb 6H4. Symbols: red squares, mBSE; blue triangles, sheep-passaged L-BSE; green circles, TgBo-passaged Sh294.