Figure 3
From: Recombinant Human Prion Protein Inhibits Prion Propagation in vitro
Inhibition of PrPSc amplification by different species of recombinant PrP.
(A) PMCA was performed with the mixture of human PrPSc (seeds) from iCJDVV2 and brain homogenates from TgWV (substrates) in the presence of different species of recombinant PrP (0.2 μM each) including mouse (Mou: rMoPrP23-231 with 129M, home-made), human (Hum: rHuPrP23-231 with 129M), bank vole (BV: rBvPrP23-231 with 109I) and bovine (Bov: rBoPrP23-231 with 129M). (B) Inhibition of PrPSc amplification was quantified using densitometric analysis based on three independent experiments. Bars represent the percentage of all five amplified PrPSc with or without inhibitors to the amplified PrPSc without any inhibitors. Amplified PrPSc was calculated by subtracting the untreated PrP intensity (−) from the PMCA-treated PrP intensity (+) shown in (A). Of all recombinant PrP species examined, recombinant human PrP23-231 exhibited the highest inhibition compared to other species (**: p < 0.01; ***: p < 0.001). (C) PMCA was performed with mouse brain homogenates infected with prion 139A (seeds) and brain homogenates from wild-type mouse FVB (substrates) in the presence of different concentrations of the commercially-derived rMoPrP23-231 with 129M. (D) The inhibition of mouse prion 139A is dose-dependent and the half maximal effective concentration (EC50) is approximately 120 nM, which is based on three independent experiments.
