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. 1996 Dec 10;93(25):14945–14949. doi: 10.1073/pnas.93.25.14945

Figure 1.

Figure 1

PrPC and PrPSc are concentrated in CLDs isolated from neuroblastoma cells using the ice-cold Triton X-100 detergent method. (A) Distribution of PrP after lysis of N2a and ScN2a cells in ice-cold Triton X-100 and separation of CLDs by flotation into sucrose gradients. Aliquots of gradient fractions were subjected to immunoblot analysis with the anti-PrP polyclonal R073 rabbit antiserum (26). Lanes: 1–7, fractions of the sucrose gradients; 8–11, lysate fractions; P, pellets. (B) Detection of PrPSc in gradient fractions. Immunoblot of gradient fractions from before (−) and after (+) treatment with PK. (C) Concentration of PrP and other proteins in CLDs. Immunoblot (Right) and silver stain (Left) of CLD proteins and cell lysate proteins from N2a and ScN2a cells. (D) Detection of PrPC degradation products in CLDs from N2a and ScN2a cells. Immunoblot of CLD proteins before (−) and after (+) treatment with PNGase F (27).