FIG. 1.

APC substrates accumulate during ΔUL97 virus infection. HFFs were mock-infected (M) or virus-infected with ΔUL97 virus (Δ) or AD169 (V) at an MOI of 3. (A) Cells were harvested over an infection time course and analyzed by Western blot assay for the expression of HCMV genes and cell cycle regulators. Actin is shown as a loading control. (B) Total RNA isolated from cells harvested over an infection time course was analyzed for geminin and G6PD expression by qRT-PCR. Geminin values were normalized to the expression of G6PD as a control for input RNA. Values are expressed as the fold induction over the level at 0 h p.i. (C) Cell samples were harvested at 16 h p.i., treated with or without (−) lambda protein phosphatase (λpp), and analyzed by Western blot assay. Equivalent cell numbers were loaded for each sample, except for lane 7 (†), which contained one-fifth the amount. Results for short and long exposures of the Cdh1 blot are shown. IE2 86 was a positive control for the phosphatase assay, while actin served as a negative and loading control. Lane numbers are shown below.