Fig 9.

γH2AX and p53 fail to accumulate in viral RCs in cells pretreated with X3 or UL54 siRNAs. HEL fibroblast cells were transfected with X3, UL54A, or CON siRNAs 24 h prior to infection with HCMV at an MOI of 0.1. Cells were fixed at indicated times p.i.; γH2AX and pUL44, or p53 and pUL44, were detected by coimmunostaining. (A) Localization of γH2AX, p53, and pUL44 proteins. γH2AX and p53 coalesce into structures reminiscent of RCs as marked by pUL44 immunostaining in control siRNA-treated cells by 48 hpi. DAPI staining is used to define nuclei. (B) Plot of the percentage of HEL fibroblasts with γH2AX or p53 foci. 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 CON, 6 hpi or 48 hpi; **, P < 0.0005 relative to CON, 72 hpi.