FIGURE 3.

Second-generation LS-PMCA. (A) Diagram of the experimental setup. Polypropylene cups were loaded with 5 mL of normal mouse BH and 5 µL of RML BH. (B) Amplification of RML prions at days 0, 1, 3, and 5 of programmed sonication in a 20 mL cup (reaction volume: 5 mL). (C) Amplification of RML prions in different 20 mL cups (reaction volume: 5 mL) from 4 separate experiments. (D) Amplification of RML prions at days 0, 1, 2, 3, and 4 of programmed sonication in a 500 mL cup (reaction volume: 100 mL). PMCA samples at different time points were treated with 50 μg/ml PK for 1 h and analyzed by SDS-PAGE and western blotting using the monoclonal antibody 6D11. For comparison, different dilutions of RML brain homogenate (2%, 5% and 10%) after PK treatment were included in the right panel.