Figure 4.

HCMV IE mRNA accumulation after infection with wt or mutant viruses in the presence of cycloheximide. (A) Human fibroblasts were infected with wt, ADsubUL82^−UL82, or ADrevUL82 virus (1 pfu per cell) in the presence of cycloheximide (100 μg/ml). At 8 h after infection, mRNA was isolated and reverse transcribed in the presence of [³²P]dCTP. The labeled cDNA was then used to probe HCMV gene arrays on membranes. Transcripts expressed after infection with the wt or revertant virus correspond to the following open reading frames: UL36–UL38, a; UL106–UL109, b; UL115–UL119, c; UL122, d; UL123, e; and actin, f. Actin was used to confirm that equal quantities of each sample were analyzed, as described in the legend for Fig. 3A. (B) Human fibroblasts were infected with wt, ADsubUL82^−UL82, or ADrevUL82 virus (1 pfu per cell) in the presence of cycloheximide (100 μg/ml). Total RNA was isolated at 8 h after infection and assayed for HCMV IE gene expression by Northern blotting with ³²P-labeled gene-specific probes.