FIG. 9.
Digital confocal images of HEp-2 cells infected with US3 null mutant virus R7037 at an MOI of 5 that were fixed and permeabilized in ice-cold methanol at 16 h.p.i. Cells were immunostained with antisera directed against UL31 and UL34 proteins. UL31 protein-specific binding (green) was detected with FITC-conjugated donkey anti-rabbit Ig antibody, and UL34 protein-specific binding (red) was detected with Texas Red-conjugated donkey anti-chicken Ig antibody. Areas of colocalization of UL31 and UL34 proteins are indicated by a yellow color upon merging of the images. Confocal Z-sections were acquired in 0.4-μm slices through the entire thickness of the cell. Digital slices from the top (row labeled Top) of the cell or the middle of the cell (row labeled Middle) are shown. A confocal Z-stacked image derived from the Z-sections is shown in the row labeled Stacked. Original magnification, X600.