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Fig. 3. Localization of huPrP, huPrP-FFI, huPrP+9OR and boPrP at the surface and in intracellular compartments of BHK cells transfected with recombinant SFV RNAs. The subcellular location was determined by immunofluorescence using the mAb anti-PrP 3B5 and a secondary Ab (goat anti-mouse IgG) conjugated with indocarbocyanine (Cy3). Cell surface immunostaining of non-permeabilized BHK cells (A-E, left panels) transfected with (A) SFV-1 RNA (without any insert), (B) recombinant SFV-huPrP1-253 RNA, (C) recombinant SFV-huPrP+90R RNA, (D) recombinant SFV-huPrP-FFI RNA and (E) recombinant SFV-boPrP1-264 RNA. (A-E, right panels) Intracellular immunostaining of permeabilized, transfected BHK cells (magnification A-E: x630).