Fig. 4
Western blot of the PK-resistant PrPSc. Brain homogenates from the frontal cortex (FC) and the cerebellum (CE) of (a) the index case, (b) GH-CJDMM1 (case 2), dCJDMM1 (case 4) and (a and b) control cases sCJDMV2-K and sCJDMM1 were treated with 10 U/ml PK (~200 μg/ml) prior immunoblotting with antibodies (i) 3 F4, (ii) Tohoku-2, or (iii) 3 F4- and Tohoku-2-merged immunoreactivities. i: The unglycosylated resPrPSc from the index case co-migrated at ~20 kDa with the corresponding resPrPSc type 1 of sCJDMM1 and with a ~20 kDa fragment of sCJDMV2-K (arrow). The asterisk in i-iii indicates the unglycosylated resPrPSc type 2. ii: Only the resPrPSc associated with sCJDMV2-K (asterisk) reacted with Tohoku-2 confirming that it belongs to type 2. Tohoku-2 immunoreacted also with a resPrPSc fragment of ~18.0 kDa and to a lower size fragment (indicated in a by the filled and empty arrowheads, respectively) in the cerebellum of sCJDMV2-K, but not with the resPrPSc ~20 kDa fragment. The dashed rectangle in ii-(a) is shown below at a longer exposure time. iii: Merged 3 F4 (green dye) and Tohoku-2 (red dye) immunoreactivities