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Fig. 1. Expression of wild-type as well as mutant human PrP and wild-type bovine PrP in transfected BHK cells and the effect of N-glycosidase F and endoglycosidase H. Immunoblot analysis of prion proteins from total cell lysates of transfected BHK cells. Cells were transiently transfected with recombinant SFV RNAs encoding wild-type human PrP (A, lanes 1 and 2; B, lanes 3 and 4), human PrP-FFI (A, lanes 3 and 4; B, lanes 5 and 6), human PrP+9OR (A, lanes 5 and 6; B, lanes 7 and 8) or wild-type bovine PrP (A, lanes 7 and 8) and with SFV-1 RNA (without any insert) (B, lanes 1 and 2). 24 hours post-transfection the expression was analyzed with mAb anti-PrP 3B5. Glycosylation patterns: di-, mono- and non-glycosylated PrP isoforms. (A) Analysis of prion proteins huPrP (lane 2), huPrP-FFI (lane 4), huPrP+9OR (lane 6) and boPrP (lane 8) after N-glycosidase F treatment. (B) Analysis of prion proteins huPrP (lane 4), huPrP-FFI (lane 6) and huPrP+9OR (lane 8) and cellular proteins from SFV-1-transfected cells (lane 2) after endoglycosidase H treatment.