FIG. 3.

Low-multiplicity infection is sufficient to deliver tegument proteins and induce the degradation of Daxx. (A) HFs were mock infected (M) or infected with HCMV at the indicated MOIs. Lysates were harvested 6 h postinfection (hpi) and analyzed by Western blotting. Tub, tubulin. (B) HFs grown on coverslips were infected with HCMV at an MOI of 0.5, 0.05, or 0.005. Cells were fixed, and delivery of pp71 to the nucleus was determined by indirect immunofluorescence. The nuclei were counterstained with Hoechst. (C) The percentage of nuclei positive for either pp71 or IE1 was compared from HCMV-infected HFs at an MOI of 0.5, 0.05, or 0.005. (D) HFs were infected at an MOI of 0.5 with gradient-purified HCMV for 4 h in the presence of cycloheximide and analyzed by indirect immunofluorescence. Fixed cells were stained for pp71 and either Daxx or PML, and nuclei were counterstained with Hoechst. Two separate panels are shown for the pp71-Daxx costaining, and a single panel is shown for the pp71-PML costaining.