FIG. 5.

Coprecipitation of Pol II and ICP8 with ICP27. HEp-2 cells were infected with either wt HSV-1 KOS1.1 or the ICP27 null mutant d27-1 virus or were mock infected. Cells were harvested at 8 h p.i. Immunoprecipitation (IP) was performed on the cell lysates with monoclonal anti-ICP27 antibody H1113. Proteins in the immunoprecipitates were resolved by SDS-PAGE and analyzed by Western blotting with monoclonal anti-Pol II antibody 8WG16 (1:1,000), monoclonal anti-ICP27 antibody H1119 (1:300), and rabbit anti-ICP8 antibody 367 (1:1,500). The ratio of loaded cell lysate to the input cell lysate for immunoprecipitation was 1:14. No Ab, wt virus-infected-cell lysate was incubated with the protein A-agarose beads in the absence of an antibody; beads, protein A-agarose beads were boiled in sample buffer.