Fig 7. The ORF2 ARM regulates the topology of ORF2 SP.

A schematic representation of differential permeabilization process with Triton X-100 and Digitonin is shown. Representative images of the differential detection of two epitopes on the ER-membrane associated Calnexin (CNX) used to assess the permeabilization conditions are shown. H7-T7-IZ cells were transfected with pTM plasmids expressing wt, mutant or chimeric ORF2 proteins. Twenty-four hours post-transfection, cells were fixed, permeabilized with either Triton X-100 or Digitonin, and processed for ORF2 staining (in red). Nuclei are in blue. Representative confocal merge ORF2/DAPI images are shown (magnification x63). Blue dots observed in some pictures are DAPI-stained transfected plasmids. A schematic representation of each construct is shown on the left and its predicted topology on the right. Blue and red asterisks correspond to 5R/5A and PSG/3R mutations, respectively. Scale bar, 20μm.