Figure 5.
In vitro prion replication by PMCA using Hsp70−/− and Hsp70+/+ brains as substrate. PMCA reactions were seeded with RML prions present in a 10−8 dilution of brain homogenate from terminally ill mice. As substrate for PMCA, we used 10% brain homogenate from three individual Hsp70−/− or Hsp70+/+ mice, and amplification continued for three consecutive rounds (see “Experimental procedures”). PrPSc formation was evaluated by Western blotting after PK digestion. Non-PK-digested samples from NBH derived from a healthy mouse was used as a migration control.