FIG. 5.
RNA synthesis of CS2 mutants. DBT cells were infected with the indicated viruses at an MOI of 5 PFU/cell as described in Materials and Methods. At 30 min prior to addition of metabolic label, medium was aspirated and replaced with prewarmed complete medium containing 20 μg/ml actinomycin D. At the indicated times postinfection, 0.03 mCi/ml of [3H]uridine was added to the medium for 2 h, and at the end of the labeling period, cells were harvested in RIPA buffer. Total actinomycin D-resistant RNA was precipitated in 10% trichloroacetic acid and quantitated by liquid scintillation counting. Data points are the means of two samples each of two concurrent independent replicate experiments for each virus.