Figure 1. Biochemical features of the protease-resistant prion protein (PrPres) detected in the brain of TgMet129, TgMet/Val129, and TgVal129 mice inoculated with vCJD. A) PrPres detected in TgMet129 (lanes 2 and 5), TgMet/Val129 (lane 3), and TgVal129 (lanes 4 and 6) mice inoculated with vCJD brain homogenate or TgMet129-passaged vCJD prions. Similar quantities of PrPres were loaded for adequate comparison and immunoblots were detected with Sha31 monoclonal antibody (mAb). The original vCJD isolate (Hu-vCJD2) used for mouse inoculations was also included in the blot (lanes 1 and 7). sCJD VV2 and MM1 isolates were included for biochemical comparative purposes (lanes 8 and 9, respectively). Molecular weight (MW) in kDa is shown. B) Glycoform analysis of PrPres from TgMet129, TgMet/Val129 and TgVal129 mice inoculated with vCJD brain homogenate or TgMet129-passaged vCJD prions. PrPres was detected by Western blot testing using the Sha31 mAb, as for panel A. The data shown are the means of >4 measurements in >2 different Western blots using the Image Lab (Bio-Rad, Hercules, CA, USA) program after capture with ChemiDoc XRS+ (Bio-Rad) under nonsaturating conditions. Error bars indicate SD. CJD, Creutzfeldt-Jakob disease; sCJD, sporadic CJD; vCJD, variant CJD.