Figure 2.

Generation of recPrP^Scby sPMCA.A, representative images of 18 rounds of sPMCA (n = 4 independent sets of sPMCA propagations). Products of sPMCA were digested with PK and analyzed by Western blotting with the 3F10 anti-PrP antibody. The seed-only reaction was a negative control, in which recPrP in the substrate was replaced by double-distilled water (ddH₂O). B, densitometric analysis of PK-resistant bands from A (n = 4). Relative convertibility = PK-resistant recPrP/recPrP without PK digestion. The relative convertibility of WT was set at 100%. C, mean convertibility of first nine rounds of sPMCA. The average convertibility WT was set as 100%. Statistical significance was determined by one-way ANOVA followed by Tukey's multiple comparison test. ∗∗∗ represents p < 0.01. Error bars indicate standard deviations. PK, proteinase K; recPrP, recombinant PrP; sPMCA, serial protein misfolding cyclic amplification.