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. 2000 May 9;97(10):5574–5579. doi: 10.1073/pnas.97.10.5574

Figure 2.

Figure 2

The amount of detergent-insoluble and protease-resistant PG14 PrP increases with age in the brains of Tg(PG14) mice. (A) Brain lysates from Tg(PG14+/+) mice of the indicated ages were subjected to ultracentrifugation, and PrP in the supernatants (S lanes) and pellets (P lanes) was analyzed by Western blotting. (B) Brain lysates from Tg(PG14+/+) mice of the indicated ages were incubated with 0–3 μg of proteinase K (PK) for 30 min at 37°C, and PrP was visualized by Western blotting. The undigested samples (0 μg/ml PK) represent 50 μg of protein, and the other samples represent 200 μg of protein. The protease-resistant fragment (PrP 27-30) migrates between 27 and 30 kDa. (C) The amount of PG14 PrP in the pellet fraction after ultracentrifugation (see A) was quantitated by densitometric analysis of Western blots of samples from Tg(PG14+/−) and Tg(PG14+/+) mice. Each bar represents the mean ± SEM of four to eight replicate analyses of samples from four to seven brains. (D) The amount of PrP 27-30 that was produced by digestion with 2 μg/ml of proteinase K (see B) was quantitated by densitometric analysis of Western blots of samples from Tg(PG14+/−) and Tg(PG14+/+) mice. Each bar represents the mean ± SEM of four to eight replicate analyses of samples from four to seven brains.