Research Paper Volume 5, Issue 3 pp 155—173

Figure 5. Determination of both PrP^Wt and PrP^Ins in detergent-soluble and -insoluble fractions by Western blotting. A and B: After ultracentrifugation in detergents, PrP in the detergent-soluble fraction (S2) (A) and -insoluble fraction (P2) (B) was detected by Western blotting with the anti-PrP antibody anti-N detecting the full-length PrP. Lane1: non-CJD control; Lane 2 fCJD^Ins+sPrPSc (case 6); Lanes 3 to 7: fCJD^Ins+rPrPSc (cases 1-5); Lane 8: sCJD control. C: Comparing compositions of total PrP, PrP^Ins, and PrP^Wt in fCJD^Ins+sPrPSc (case 6) and fCJD^Ins+rPrPSc (cases 1-5, mean) by densitometric analysis of PrP intensity detected with Western blotting as shown in A and B. The left four bars exhibit the percentage of PrP^Wt or PrP^Ins in total PrP including soluble and insoluble forms. The central four bars exhibit the percentage of soluble or insoluble form in total PrP^Ins. The right four bars exhibit the percentage of soluble or insoluble form in total PrP^Wt. There were no significant differences in the percentage of PrP^Wt or PrP^Ins in total PrP between fCJD^Ins+sPrPSc and fCJD^Ins+rPrPSc (62.5/37.5 vs 63.4/36.6). Also there were no significant differences in the percentage of the soluble or insoluble form in PrP^Ins between the two conditions (36.2/63.8 vs 30.5/69.5). In contrast, the percentage of the soluble form of PrP^Wt was remarkably greater in fCJD^Ins+sPrPSc than in fCJD^Ins+rPrP (54.7% vs 38.4%), while the percentage of the insoluble form of PrP^Wt was significantly smaller in fCJD^Ins+sPrPSc than in fCJD^Ins+rPrPSc (45.3% vs 61.6%). D: Comparison of PrP^Wt intensity in S2 and P2 fractions between fCJD^Ins and non-CJD or sCJD. No differences in the levels of PrP^Wt from soluble or insoluble fraction were detected between fCJD^Ins and non-CJD or sCJD.