FIG. 5.

μNS aa 75 to 84 are necessary and aa 55 to 173 are sufficient for associations with the core surface protein λ2. For each experiment, cells were processed for fluorescence microscopy at 18 h p.t. (A) CV-1 cells were cotransfected with plasmids expressing λ2 and either μNS (top row), μNS(55-721) (second row), μNS(65-721) (third row), μNS(75-721) (fourth row), μNS(85-721) (fifth row), or μNS(95-721) (bottom row). After fixation, cells were stained with rabbit polyclonal antibodies against μNS and mouse monoclonal antibody 7F4 against λ2 followed by Alexa 594-conjugated goat anti-rabbit IgG to visualize μNS (left) and Alexa 488-conjugated goat anti-mouse IgG to visualize λ2 (right). (B) CV-1 cells were cotransfected with plasmids expressing λ2 and either EGFP/NSP5 (top row), μNS(1-227)/EGFP/NSP5 (second row), μNS(41-221)/EGFP/NSP5 (third row), μNS(55-221)/EGFP/NSP5 (fourth row), or μNS(95-221)/EGFP/NSP5 (bottom row). After fixation, cells were stained with mouse monoclonal antibody 7F4 against λ2 followed by Alexa 594-conjugated goat anti-mouse IgG to visualize λ2 (right). The inherent fluorescence of EGFP was used to visualize each of the fusion proteins (left). (C) CV-1 cells were cotransfected with plasmids expressing λ2 and either μNS(55-173)/EGFP/NSP5 (top), μNS(95-173)/EGFP/NSP5 (middle), or μNS(41-110)/EGFP/NSP5 (bottom). After fixation, cells were stained with mouse monoclonal antibody 7F4 against λ2 followed by Alexa 594-conjugated goat anti-mouse IgG to visualize λ2 (right). The inherent fluorescence of EGFP was used to visualize each of the fusion proteins (left). Bar, 10 μm.