Fig 2.

Expression of HCMV pUL29/28 and pUL38 alters p53-regulatable gene expression in the absence of infection. (A) Control U2OS or U2OS-pUL38 (+pUL38) were transfected with a plasmid containing the p21CIP1 promoter driving luciferase along with 1.0, 10.0, and 100.0 ng of either empty pCGN or pUL29/28 plasmid DNA. Relative luciferase activity was measured at 48 h after transfection. The data are means for two replicate samples ± standard deviations. Statistical analysis was performed using Student's t test (**, P ≤ 0.01). (B) The U2OS and U2OS-pUL38 (+pUL38) cells were either transfected with empty vector or pUL29/28, or treated with nutlin-3 for 24 h. Cells were harvested 48 h posttransfection. Western blot analysis was performed using antibodies against the indicated proteins. (C) U2OS cells were transfected with pUL29/28HA expression vector. At 40 h posttransfection, the cells were treated with or without nocodazole to synchronize the population. Cells were stained using antibody against the HA epitope and propidium iodide, and DNA content was measured by flow cytometry. The data are displayed as percentages of cells in each phase of cell cycle and are the means from two biological experiments ± standard errors of the means. Statistical analysis was performed using Student's t test (*, P ≤ 0.05; **, P ≤ 0.01).