FIG 4.
Highly efficient amplification of murine, hamster, and human prions by a single round of mb-PMCA. Western blot analysis (a) and ratio (b) of high- and low-molecular-mass PrPres glycoforms of mb-PMCA products. Brain homogenates from tga20 mice infected with Chandler prions, tg7 mice infected with 263K prions, and tg650 mice infected with vCJD prions were serially diluted in homotypic brain lysates as indicated. Each dilution was directly analyzed by Western blotting or was submitted to a single round of mb-PMCA, as indicated (a). All samples were treated with PK before SDS-PAGE and Western blotting. Data in panel b are plotted as means ± standard errors of the means. Nonamplified, brain prions are represented by red triangles; mb-PMCA amplicons generated from the indicated input seed are indicated by blue symbols. The ratios were established from 3 independent mb-PMCA experiments.