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. 2013 Jun 13;22(21):4253–4266. doi: 10.1093/hmg/ddt276

Figure 4.

Figure 4.

PrP-α3M>S mutants acquire Ctm topology. (A and B) Tissue homogenates from flies expressing PrP-WT, -M205S and -M205,212S subjected to a PK gradient at Days 1 (A) and 30 (B). PrP-WT is degraded by the lowest concentration of PK regardless of age. PrP-M205S and -M205,212S are partially resistant to PK, which increases over time. (C) Protease protection in microsomes. In the absence of a detergent, microsomes protect full-length PrP-WT from PK in 1-day-old flies, but 30-day-old flies also accumulated Ntm and Ctm PrP. In young PrP-M205S and -M205,212S flies, microsomes reveal full-length and Ctm PrP, which increases in older flies. In the presence of detergent, PK degrades all PrP. (D) The SP antibody detects the two mutants (bottom), whereas 6D11 detects all PrPs in the same membrane (top) (E) Model of secreted and Ctm PrP indicating the TMD and the SP.