FIG. 2.

The level of Wee-1 remains stable following Vpr-induced arrest. (A) HeLa cells (2.0 × 10⁶) were synchronized at G₁/S by a double thymidine block. They were then infected with equivalent amounts of HR′Vpr or HR′EGFP (4.0 μg of viral p24). Five hours after infection cells were subjected to a 30-min period of serum starvation and pulse-labeled for 10 min with ³⁵S-labeled methionine-cysteine. This was followed by a chase period for 5 h. Cells were analyzed during the chase period at the time points indicated. Equivalent amounts of cell lysates were immunoprecipitated with antibody specific for Wee-1. Antibody-bound immune complexes were separated by SDS-10% PAGE. ³⁵S-labeled Wee-1 was detected by autoradiography. Data shown are representative of results from three independent experiments. (B) Quantitation of Wee-1 levels by densitometry. The signal obtained at the starting point (0 h) for HR′EGFP virus-infected HeLa cells was arbitrarily set to 100%. P.I., postinfection.