FIG 7.

US3 prevents p65 nuclear translocation. HeLa cells were transfected with control vector, US3-Flag, K220M-Flag, and D305A-Flag expression plasmids. Twenty-four hours posttransfection, cells were treated with TNF-α or mock treated for 30 min as indicated. Cells were stained with mouse anti-Flag and rabbit anti-p65 pAb. FITC-conjugated goat anti-rabbit (green) and TRITC-conjugated goat anti-mouse (red) were used as the secondary antibodies. Cell nuclei (blue) were stained with Hoechst 33258. The images were obtained by fluorescence microscopy using a 40× objective.