Immunoblots of PrP^Sc in cerebellar slice cultures infected with RML strain.

<p>A) The typical banding pattern of RML-infected brain homogenate (RML BH) inoculum after proteinase K (PK) digestion is shown. 10, 5, and 2 µg total protein were loaded. B) RML-infected POSCA slices were homogenized in pairs in RIPA buffer, then 30 µg total protein was treated with 20 µg/mL PK for one hour (+). Undigested slice culture homogenates (−) were loaded at 10x lower concentration for comparison. PrP^Sc is distinguished by its truncated state after PK digestion, seen as lower molecular weight bands when compared with PrP^C. PrP^Sc can also been seen to increase in amount with increasing culture time (days 7 through 42 are shown). C) RML-treated POSCA slices from <i>Prnp^0/0</i> mice did not show PrP^Sc at any time point. Day 45 is shown.