FIG 5.
Transient transfection of UL13-PK but not of US3-PK stabilized ICP0 at early times after infection. (A) Schematic representation of experimental design. HEK293T cells were transfected with plasmids encoding UL13-PK or US3-PK 24 h before infection. The protocols for infection, the replacement of medium after 1 h, the addition of cycloheximide at 3 h after infection, and analyses of cells harvested at 3, 6, or 10 h after infection were similar to those indicated in legends to Fig. 1 to 3. (B) Immunoblots of cells infected with HSV-1(F). Lanes 1 and 2, uninfected mock-treated or pcDNA3-transfected cells, respectively; lanes 3 to 14, HSV-1(F)-infected mock treated cells (lanes 3 to 5), or cells that had been transfected with pcDNA3, or plasmids encoding UL13 or US3 protein kinases as indicated. (C) Same as panel B except that the cells were infected with the ΔUL13 mutant virus. In both panels the electrophoretically separated proteins transferred to a nitrocellulose membrane were reacted with the antibodies shown.