FIG 2 .

BADinUL71STOP produces a higher proportion of cell-associated progeny than BADwt but accumulates normal levels of viral DNA and proteins. Infections were performed at a multiplicity of 2 PFU/cell. (A) BADinUL71STOP produces >100-fold-fewer extracellular infectious viral progeny and ~10-fold-fewer cell-associated infectious viral progeny than BADwt. Cultures were harvested on the indicated days postinfection (dpi), and viral titers were determined by TCID₅₀ assay. Inp, input virus. Results are representative of an experiment conducted in duplicate with two separate isolates of BADinUL71STOP mutant virus. Error bars estimate standard deviations calculated from technical repeats. (B) Ratio of cell-associated to cell-free virus for mutant and wild-type viruses. The primary data displayed in panel A are plotted as ratios. Error bars represent standard errors of means calculated from technical repeats. (C) BADinUL71STOP-infected cells accumulate viral DNA normally. Fibroblasts were harvested at the indicated hour postinfection (hpi) with mutant or wild-type virus, and total cellular DNA was isolated. Viral genomes were quantified by qPCR with UL123-specific primers and normalized to cellular actin. Results are representative of an experiment conducted in duplicate with two separate isolates of BADinUL71STOP. Error bars represent standard errors of means calculated from technical repeats. (D) BADinUL71STOP-infected cells accumulate representative viral proteins from all kinetic classes normally. Cells were harvested at the indicated hpi, and whole-cell lysates were prepared. Proteins were analyzed by Western blotting using antibodies for IE1, pUL44, and pUL99. Tubulin was assayed as a loading control. Results are representative of an experiment conducted in duplicate with two separate isolates of BADinUL71STOP.