Fig 5.

HSV-2 ΔUL21 displays a delay in virus protein expression. (A) Vero cells were infected with HSV-2 WT, ΔUL21, or ΔUL21R at an MOI of 0.1. At the indicated times postinfection, cells were fixed and analyzed for ICP0/ICP8 (IE/E) and ICP0/gD (IE/L). Cells were stained with rat polyclonal antiserum specific for ICP0 and mouse monoclonal antibodies against ICP8 or gD, followed by staining with Alexa Fluor 568-conjugated goat anti-rat IgG (red signal) and Alexa Fluor 488-conjugated donkey anti-mouse IgG (green signal). Nuclei were visualized with Hoechst 33342 (blue signal). Images were captured by using confocal microscopy. Representative images are shown. Scale bars, 10 μm. (B) Vero cells were infected with HSV-2 WT, ΔUL21, or ΔUL21R at an MOI of 1.0. At the indicated times postinfection, cells lysates were analyzed by immunoblotting against representative proteins from three kinetic classes: IE (ICP27), E (ICP8 and Us3), and L (VP16, ICP5, and gD). Molecular mass markers in kDa are shown on the left.