FIG. 6.

The canonical NF-κB pathway is not required for replication of human CMV in fibroblasts. (A) NHDF were transduced with control (LXSN) or IκBαM retroviral vector. Cells were examined for upregulation of ICAM-1 by flow cytometry after incubation with ±1 nM TNF. Dotted histogram represents binding of isotype control antibody. (B) Nuclear extracts from NHDF-LXSN or -IκBαM (+) cells treated with TNF (1 nM) or LTα (1 nM) or infected with HCMV or UV light-inactivated HCMV (uvHCMV; MOI, 5 or 2) for 2 h were subjected to EMSA to analyze the level of p50/p65 NF-κB (indicated by the arrow) translocation to the nucleus. (C) NHDF-LXSN or -IκBαM cells were infected at an MOI of 0.01 or 2 with HCMV strain AD169 or Toledo. Cell supernatant was collected at various days postinfection, and viral titers (PFU/ml) were determined in triplicate by plaque assays in NHDF. Error bars represent standard errors of the means.