Radioactivity was detected either by autoradiography or using a phosphorimager (Typhoon, GE Healthcare).
Sequence exchanges were detected either visually or using the statistical program for detecting gene conversions GENECONV version 1.18 [ 33].
Immunosignals were detected either directly under epifluorescence microscopy or after using a Vectastain Elite ABC kit (Vector Laboratories) with 3,3′-diaminobenzidine visualization (Roche Applied Science, Indianapolis, IN).
The antibody was detected using either peroxidase conjugated goat anti-human IgG or IgM (secondary antibody Caltag – H10007, H 15007).
Cells were lysed at 24 hours after transfection, normalized for protein concentration, and following SDS-PAGE and immunoblotting, NPM1NS1 and NPM1M2NS1 expression was detected using either anti-HA-tag or anti-Flag antibodies (Cell Signaling, Beverly, MA, USA).
In the ascitic fluid, the light and heavy chains are thus secreted with a correct 1∶1 stoichiometry and are all assembled into Ig molecules since no fragment other than the two chains was detected using either non-reducing or reducing conditions (Figure 2A, lane 1 and figure 2B, lanes 3 and 4).
PrP was detected using either monoclonal mAb 3F4 or monoclonal mAb IPC1 (Sigma-Aldrich Corp .. Protein bands were visualized with an anti-mouse HRP secondary antibody (Pierce Biotechnology Inc., Rockford, Illinois, United States) and enhanced chemiluminescent detection (Pierce Biotechnology Inc .. Hamster brains were fixed in 10% neutral buffered formalin for 48 h then sunk in 30% sucrose.
Signal on the blot was detected using either myECL Imager (Thermo Scientific) or a Storm 865 (GE Healthcare).
V5 epitope tag was detected using either mouse anti-V5 mAb (Life Technologies, Grand Island, NY) or a rabbit pAb (EMD Millipore).
The expression of each protein was detected using either FITC, PE or Cy3 conjugated secondary antibodies (at 1 300 dilution).
