Destabilizing polymorphism in cervid prion protein hydrophobic core determines prion conformation and conversion efficiency
(a) The curves depict a representative RT-QuIC response of serially diluted (2x10-2 to 2x10-7) wt (left panels) and 116AG (right panels) brain homogenates using rPrP deer or mouse substrate. Fluorescence signals were measured every 15 min. The x-axis represents the reaction time (hours) and the y-axis represents the relative fluorescence units (RFUs), and each curve represents a different dilution. Mean values of four replicates were used for each dilution. The cut-off is indicated at app. 50,000 RFU based on the average fluorescence values of negative control +5SD. (b, c) The RT-QuIC responses of wt and 116AG were quantified by calculating different parameters: lag phase (b), and log phase (c). Mean values of 5 experiments with 4 replicates each were used and statistical analysis were evaluated using log-rank (Mantel-Cox) test for the lag phase (b) and unpaired t-test for the log phase (c). *P <0.05, **P <0.01 and ***P <0.005 refers to differences between WTD isolates (GraphPad Prism software).