Fig. 2.

Q548R IE2 C-F HB5 virus is capable of genome replication and geminin stabilization. G₀-synchronized HFFs were infected at the time of release into G₁ with either WT C-F HB5, Q548R IE2 C-F HB5, or Rev-Q548R IE2 C-F HB5 (MOI = 1) or mock infected. Cells were harvested at the indicated times. (A) DNA from the 24-, 72-, and 96-h p.i. samples was isolated and analyzed via quantitative PCR of the UL77 gene. UL77 levels were normalized to levels of the cellular glyceraldehyde 3-phosphate dehydrogenase promoter in each sample. Values are relative amplification (Amp), with the lowest value set to 1. The experiment was carried out three times, and representative results are displayed. (B) Protein was isolated from the 24-, 48-, 72-, and 96-h p.i. samples. Equivalent amounts of protein were loaded on an 8% acrylamide gel and transferred to a nitrocellulose membrane. The membrane was then probed for geminin, and β-actin served as a loading control.