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. 2012 Mar;86(5):2523–2532. doi: 10.1128/JVI.06548-11

Fig 5.

Fig 5

Characterization of UL94stop phenotype. (A) The UL94stop virus is completely defective for replication. HFF cells were infected with wild-type virus (●), UL94HA virus (○), UL94 stop virus (▼), or UL94 repair virus (▽) at a multiplicity of 0.01 PFU/cell. Total virus (cells and supernatant) was harvested at the indicated days postinfection. Harvested virus titers were determined by plaque assay on HAUL94 complementing cells. (B) Expression of viral proteins over a time course of infection. HFF cells were infected with UL94HA virus or UL94stop virus at a multiplicity of 0.01 PFU/cell, and total protein was harvested at the indicated times postinfection. Viral proteins were analyzed by Western blotting with the indicated antibodies. (C) Quantitative PCR analysis of viral genome replication over time. HFF cells were infected with UL94HA virus (○) or UL94stop virus (●) at a multiplicity of 0.01 PFU/cell, and total DNA was harvested at the indicated times postinfection. Accumulation of viral DNA was analyzed by quantitative PCR. The results were normalized for both input DNA and variations in DNA recovery introduced during purification. Picograms of viral DNA present in each sample were calculated from a standard curve prepared from purified viral DNA of a known concentration.