FIG. 1.

Differentiation of cells newly infected with VZV from the infected-cell inoculum. The labeling method and selective analysis of newly infected (output) cells are outlined in the upper left panel. Examples of the spatiotemporal expression patterns of several viral proteins in newly infected cells (white stars) are shown. Unlabeled and uninfected (output) HELF cells were seeded on coverslips and infected with green-CellTracker-labeled and infected inoculum cells. The cells were then fixed at various time points and stained with specific antibodies for VZV proteins (red) IE62 and ORF61 (fixation at 4 h), IE63 and ORF29 (fixation at 6 h), or gE and ORF23 (fixation at 9 h) and Texas Red-conjugated secondary antibodies. Viral DNA (vDNA; red) was detected by DNA in situ hybridization (fixation at 6 h). The cell nuclei (blue) were counterstained with Hoechst 22358. The white arrow in the panel for ORF23 marks a newly infected cell nucleus that is also shown at higher magnification (inset). Scale bars are 20 μm.