FIG 6.

In trans expression of gpUL132 rescues the assembly compartment formation phenotype in ΔUL132 HCMV-infected cells. (A and B) Transient expression of gpUL132 rescues the AC formation phenotype in ΔUL132 HCMV-infected cells. Confluent HFFs were mock treated or electroporated with a vector expressing UL132 and then infected with WT or ΔUL132 HCMV. Infected cells were harvested at 4 dpi, fixed, processed for immunofluorescence staining of gB and pp65, and analyzed using confocal microscopy. (A) Transient expression of gpUL132 rescues the AC in ΔUL132 HCMV-infected cells. Representative images are shown. The images in the top two rows illustrate the morphology of the AC as demarcated by gB and pp65 staining in WT and ΔUL132 HCMV-infected cells compared to the cells electroporated with a vector expressing UL132 and then infected with the respective viruses in the bottom panels. Magnification, ×120. (B) Quantification of AC formation in transiently expressed gpUL132-infected HFFs. (C and D) The WT AC phenotype is rescued in ΔUL132 HCMV-infected UL132-expressing cells. Confluent UL132-expressing HFFs were infected with WT or ΔUL132 HCMV. Infected cells were harvested at 4 dpi; fixed; processed for immunofluorescence detection of gB, pp65, and HA-tagged UL132; and analyzed using confocal microscopy. (C) Representative images of AC formation in UL132-expressing cells. Magnification, ×120. (D) Quantification of AC formation in UL132-expressing HFFs during infection. ACs that formed in nine random fields with 35 to 45 cells per field were counted, and the percentage of ACs per field was plotted. A total of 349 WT HCMV-infected cells and 320 ΔUL132 HCMV-infected cells were counted. Error bars denote standard deviations (SD) of the means. Significance was determined using the Student t test (*, P < 0.05; ns, not significant). The experiments were repeated twice, and the data shown are from one experiment.