Full restoration of specific infectivity and strain properties from pure mammalian prion protein
Fig 2
Protein-only recPrPSc seeds effectively propagate in BV BH, but not Mo BH sPMCA reactions.
Western blots probed with anti-PrP mAb 27/33. (A) Titration of Mo protein-only recPrPSc in vitro. Western blots showing three-round BH sPMCA reactions with either Mo BH (top row) or BV BH (bottom row) substrates. Reactions were seeded with ten-fold serial dilutions of Mo protein-only recPrPSc. The 10−1 reaction was seeded with 6 μg/mL of Mo protein-only recPrPSc for a final reaction concentration of 0.6 μg/mL of seed. (B) Titration of BV recPrPSc seeds in vitro. Western blots showing the third round of three-round BH sPMCA reactions with either BV BH (left column) or Mo BH (right column) substrates. Reactions were seeded with 10-fold serial dilutions of the indicated BV recPrPSc seed. The 10−1 reactions were seeded with 6 μg/mL of recPrPSc for a final reaction concentration of 0.6 μg/mL. NS- no seed. (C) M109 recPrP amyloid seeded three-round BV BH sPMCA reactions. Reactions were seeded with the increasing concentrations of amyloid, as indicated. Note that input recPrP amyloid seeds migrate at a lower MW than the converted native PrPSc sPMCA product, as indicated by the boxed labels.