FIG. 6.

MHC class I is redistributed to the Golgi region in the presence of CVB3 2BC. (A) EGFP(mem)-expressing dicistronic constructs were transfected into HeLa (RW) cell monolayers, and at 16 h posttransfection, MHC class I (HLA-ABC) (red fluorescence, first column) and nuclei (blue fluorescence, third column) were stained. In contrast to the approach taken in Fig. 4, the cells depicted here were subjected to a permeabilization step prior to HLA-ABC staining to reveal not only surface but also intracellular MHC class I localization. Confocal z-series images were acquired, and z-dimensional profiles, shown below each image, were generated along the x-y axes indicated by the yellow lines. A and B, the apical and basal sides of the cell monolayers, respectively. Merged images for each of the four constructs tested are shown in the last column. (B) To assess the localization of intracellular MHC class I relative to the Golgi complex in cells expressing 2BC, cells transfected with this construct were stained using a rabbit polyclonal antibody to giantin, a major component of the Golgi complex (fourth column; depicted as green fluorescence in the merged image). HLA-ABC staining is depicted in red as described above, and nuclei were revealed by staining with DAPI. The merged image shows colocalization between MHC class I and the Golgi subcompartment.