GPI^–RML prions inoculated into WT mice (GPIRML-WT).

<p>(A) Survival curves of WT mice inoculated with RML or GPI^–RML prions (passage 2). (B) PrP and astrocyte immunostaining of brain sections at the level of cerebral cortex and hippocampus show diffuse PrP^Sc deposits and accompanying gliosis. (C) Lesion profile analysis. For RML- and GPIRML-infected WT mice, the severity of spongiosis, astrogliosis, and PrP^Sc deposition were scored for nine brain regions (see Methods) and were nearly superimposable. Each ring represents 1 point. (D) PK-digested brain samples from WT mice infected with RML or GPI^–RML show no difference in their glycoform profiles. Also shown are Tg(GPI^–RML) mice infected with RML, which show a shift to a lower molecular weight and only mono-glycosylated PrP. (E) Conformational stability and (F) thermal stability of RML and GPIRML-WT were nearly identical (n = 4 mice each). (G) Thermal denaturation curves of RML- and GPIRML-infected WT mice show the insoluble PrP^Sc remaining after heating to various temperatures as measured by ELISA. Right panel, T_1/2 values were independently calculated for each mouse and plotted. For (E), plotted are the averages from n = 4 mice per prion, each run in triplicate. For (F), the middle graph shows the mean ± SE of RML and GPI^–RML (n = 4 mice each). Graphs (right panels) represent the mean ± SE for all mice. Scale bar = 500 µm.