Table 2.
Automated serial PMCA using WT or Tg(MoPrP167S) brain homogenate as the substrate
| PrPC source | Experiment 1 (4 rounds) | Experiment 2 (5 rounds) |
|---|---|---|
| Unseeded | ||
| WT | 0/3 | 0/3 |
| Tg(MoPrP167S) | 0/3 | 0/3 |
| Sheep (SSBP1) | ||
| WT | 0/3 | 0/3 |
| Tg(MoPrP167S) | 0/3 | 0/3 |
| Deer CWD | ||
| WT | 0/3 | 0/3 |
| Tg(MoPrP167S) | 0/3 | 0/3 |
| Hamster (263K) | ||
| WT | 0/3 | 0/3 |
| Tg(MoPrP167S) | 0/3 | 0/3 |
| Mouse (RML) | ||
| WT | 3/3 | 3/3 |
| Tg(MoPrP167S) | 3/3 | 3/3 |
| Cattle BSE | ||
| WT | 1/3 | 3/3 |
| Tg(MoPrP167S) | 1/3 | 2/3 |
Brains from WT and Tg(MoPrP167S) mice were extracted after perfusion with PBS + 5 mM EDTA and used as substrate or source of PrPC. Brain homogenates (60 μl) were treated in replicates of 3 (n=3) with saPMCA and were incubated-sonicated for 27 h in each round. After each round of PMCA, the PK-resistant PrP (PrPSc) in the incubated-sonicated samples was diluted 1:10 into a freshly prepared WT and Tg(MoPrP167S) mouse brain homogenate to start the next PMCA round. This process was repeated for 4 or 5 rounds (P5). The fraction of PrPSc-positive tubes is indicated.