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. 2009 Nov 4;84(2):867–882. doi: 10.1128/JVI.01571-09

FIG. 4.

FIG. 4.

μNS aa 65 to 74 are necessary and aa 41 to 173 are sufficient for associations with the core surface protein λ1. For each experiment, cells were processed for fluorescence microscopy at 18 h p.t. (A) CV-1 cells were cotransfected with plasmids expressing λ1 and either μNS (top row), μNS(55-721) (second row), μNS(65-721) (third row), μNS(75-721) (fourth row), μNS(95-721) (fifth row), or μNS(173-721) (bottom row). After fixation, cells were stained with rabbit polyclonal antibodies against MRV cores followed by Texas Red-conjugated μNS-specific rabbit IgG to visualize μNS (left) and Alexa 488-conjugated goat anti-rabbit IgG to visualize λ1 (right). (B) CV-1 cells were cotransfected with plasmids expressing λ1 and either EGFP/NSP5 (top row), μNS(1-227)/EGFP/NSP5 (second row), μNS(41-221)/EGFP/NSP5 (third row), or μNS(55-221)/EGFP/NSP5 (bottom row). After fixation, cells were stained with rabbit polyclonal antibodies against MRV cores followed by Alexa 594-conjugated goat anti-rabbit IgG to visualize λ1 (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 λ1 and either μNS(41-173)/EGFP/NSP5 (top) or μNS(41-110)/EGFP/NSP5 (bottom). After fixation, cells were stained with rabbit polyclonal antibodies against MRV cores followed by Alexa 594-conjugated goat anti-rabbit IgG to visualize λ1 (right). The inherent fluorescence of EGFP was used to visualize each of the fusion proteins (left). Bar, 10 μm.