FIG. 5.
GdnHCl-induced denaturation of PrPSc from the cerebellums and thalami of vCJD cases. (A) Extracts of cerebellums and thalami from vCJD cases were centrifuged, and the resultant pellets were exposed to increasing concentrations of GdnHCl. The degree of unfolding of PrPSc was measured by CDI, and the denatured fraction of PrPSc was calculated (empty squares, cerebellum; filled squares, thalamus). Data shown represent the average values ± SD for three individuals. The result for the cerebellum was an average value from three independent experiments in triplicate, and the result for the thalamus was an average value from a one-time experiment in triplicate. (B) Based on denaturation profiles shown in panel A and Fig. 3, the GdnHCl1/2 values were interpolated using the best fit of a second-order polynomial curve in Excel. Each symbol represents the value of an individual sample (vCJD, gray; sCJD, black). The results for three vCJD cases (diamonds, vCJD1; squares, vCJD2; triangles, vCJD3) and four sCJD cases (diamonds, MM1-1; squares, MM1-2; triangles, MM1-3; circles, VV2) are shown. The GdnHCl1/2 values of five samples (three FC samples, one cerebellum [Cb] sample, and one thalamus [Th] sample) in each vCJD case, two FC samples in each case of sCJD MM1 subtype, and one FC sample of a sCJD subtype VV2 case are depicted.