FIG. 1.
Vpr stabilizes Wee-1 in Vpr-infected HeLa cells. HeLa cells (1.0 × 106) were synchronized at G1/S by a double thymidine block and then infected with equivalent amounts (2.0 μg of viral p24) of HR′Vpr or HR′EGFP. Infected cells were cultured for various times as indicated prior to analysis. (A) Cells were stained with propidium iodide and analyzed with a FACScan cell sorter. A total of 10,000 events were collected and analyzed in each sample. (B) Whole-cell lysate (10 μg) was separated by SDS-7.5% (Wee-1) or 15% (others) PAGE and probed with antibodies to Wee-1, phosphorylated Cdc2, total Cdc2, HA-tagged Vpr, and actin (as a loading control). Data shown are representative of Western blotting results obtained in three independent experiments.