FIGURE 4.
Characterization of PrPC substrate. A, gray (G) and white (W) matter-enriched homogenates prepared from each PrP genotype showed increased NeuN and MBP reactivity, respectively, and similar levels of β-actin. B, the ratio of PrP expression in white:gray-enriched brain indicated that there was more PrP present in gray matter-enriched brain from MM and MV PrPC substrates, but unchanged in the VV PrPC substrate. C, the proportions (%) of di- (triangles), mono- (circles), and unglycoslated (diamonds) forms of PrPC were calculated from the Western immunoblot (mean ± S.E. (error bars)). D, Western immunoblotting was performed of homogenates enriched for gray and white matter from MM, MV, and VV PrPC substrates treated with (+) or without (−) peptide:N-glycosidase F and probed with 3F4. E, full-length PrP (arrow) and truncated PrP (bracket) were quantified and are shown as the percentage full-length of total. For each analysis n = 4 (MM), n = 5 (MV), n = 3 (VV). Significance (*, p < 0.05; **, p < 0.01; ***, p < 0.001; t test) was calculated for each glycotype. Error bars, S.E.