WB analysis of PrPSc digestion in different partially
denaturing conditions. A, immunoblot analysis of PrPSc
from frontal cortex of all sCJD subtypes and vCJD. Samples were incubated in
GdnHCl 1 m for 1 h and untreated or treated with PK. B,
immunoblot analysis of PrPSc from frontal cortex of an MM1 case
digested with PK at different temperatures. The incubation time was 30 min for
all samples but one (marked with an asterisk), which was incubated
for 1 h. C, immunoblot analysis of PrPSc from frontal
cortex of an MM1 case. Standard (i.e. “PMSF+”) and
“PMSF–” conditions, as indicated under “Experimental
Procedures,” are compared. Membranes were probed by the 2301 antiserum.
D, immunoblot analysis of PrPSc from frontal cortex of an
MM1 and a VV1 subject. All samples are in PMSF–conditions and
deglycosylated. Membranes were probed by the 2301 antiserum. The 16-kDa
fragment generated in denaturing conditions (DCF 16) is marked with an
asterisk. All results were reproduced twice with samples from nine
sCJDMM1/MV1 and at least three subjects for each of the other groups.
Approximate molecular masses are in kilodaltons.