FIG. 6.

NF-κB inhibitory drugs block IE gene expression. Cells were treated with aspirin or MG-132 for 1 h prior to infection with HCMV (MOI, 3 to 5). (A) Protein levels were analyzed by Western blot analysis using antibodies specific for IE1-72. Replicate studies were performed using IE2-86-specific reagents with similar results (data not shown). Equal protein was loaded per lane. (B) Steady-state mRNA levels were analyzed by RT-PCR analysis. RT-PCR was performed using primers specific for IE1-72. Equal RNA loading was determined by repeating the PCR using GAPDH-specific primers. Replicate studies were performed using IE2-86-specific reagents with similar results (data not shown). Fibroblasts treated with the drug solvents (1 M Tris [pH 8.0] for aspirin and dimethyl sulfoxide for MG-132) and infected with HCMV were used as controls, which showed no effect due to the solvents (data not shown). Experiments were repeated with similar results.