Fig 7.

Pretreatment with X3 and UL54 siRNAs alters the formation of viral replication compartments (RCs). HEL fibroblast cells were untransfected or transfected with X3, UL54A, or CON siRNAs 24 h prior to infection with HCMV at an MOI of 1.0. Cells were fixed at 48 hpi, and pUL44 and IE proteins were detected by immunostaining. (A) Localization of IE and pUL44 proteins. Cells with “immature” RCs were defined as those with multiple, small pUL44 compartments (for example, see the yellow arrow), and cells with “mature” RCs were identified as those composed of single, larger pUL44 compartments typically representing more than two-thirds of the nucleus (for example, see the white arrow). 4′,6-Diamidino-2-phenylindole (DAPI) staining is used to define nuclei. (B) The percentage of fibroblasts with mature RCs was plotted relative to the percentage of those lacking RCs or having immature RCs. The percentage of fibroblasts with IE staining was also determined. Over 200 cells were scored per sample. Histograms show the averages of three independent experiments, and the error bars denote the standard deviations.*, P < 0.005 relative to X3; **, P < 0.0005 relative to X3; ***, P < 0.0001 relative to X3.