FIG 5.

All the mutated rabbit rec-PrP^res constructs induce prion-like misfolding of wild-type rabbit rec-PrP in vitro. (A) Representation of the mean (± standard deviation) of the maximum dilution reached by each mutated rec-PrP^res-based seed on a substrate containing wild-type rabbit rec-PrP. The x axis shows the different mutated rec-PrP^res constructs acting as seeds, grouped according to their origins (recombinant [for those misfolded by the recRML seed] or brain [for the ones misfolded using brain-derived RML as a seed]). (B) Western blot representing PK-digested (85 μg/ml) misfolded wild-type rabbit rec-PrP^res. A wild-type rabbit rec-PrP-based substrate was seeded with 11 different mutated seeds and subjected to 15 rounds of recPMCA. Similar bands of approximately 17 kDa, corresponding to the PK-resistant 90-230 fragment of the wild-type protein, are shown. Membranes were developed with monoclonal antibody SAF84 (1:400). Rabbit recPrP, undigested recombinant rabbit rec-PrP.