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Fig. 2. Transduction of Prnp–/– cells with lentivirus carrying the PrP dominant negative mutants MoPrPQ167R and MoPrPQ218K. (A) Transduction efficacy of Prnp–/– cells with lentiviral vectors assessed by GFP fluorescence. Images A1, A2, A3 and A4 correspond to non-transduced, GFP, MoPrPQ167R and MoPrPQ218K transduced cells, respectively. (B) Detection of PrPC by immunoblotting using SAF32 anti-PrP antibody (C) SAF32 immunocytofluorescence of the PrPC in Prnp–/– cells: no staining is visible in (C5) non-transduced and (C6) GFP lentiviral transduced cells, whereas cells transduced with the (C7) MoPrPQ167R, or (C8) MoPrPQ218K lentivirus exhibit a strong rhodamine fluorescence. C1, C2, C3, C4 correspond to the Hoechst nuclear staining of the cells represented in figure C5, C6, C7, C8 respectively. Scale bars, 8 µm.





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