Fig 2.

UL99 interacts with and stabilizes UL94 protein. (A) 293T cells were transfected with HAUL94 alone or with FLAGUL99. Total protein was harvested at 48 h posttransfection, and immunoprecipitation (IP) was performed with anti-FLAG antibody. Immune complexes were separated by SDS-PAGE, and Western blotting (WB) was performed with anti-HA antibody. (B) 293T cells were cotransfected with the indicated amounts of plasmid DNA expressing HAUL94 and FLAGUL99. Total protein was harvested at 48 h posttransfection, and levels of UL94 and UL99 protein were analyzed by Western blotting. (C) 293T cells were transfected with plasmid DNA expressing HAUL94 alone or cotransfected with FLAGUL99, as indicated. Cells were treated with cycloheximide (CHX) at 48 h posttransfection. Total protein was harvested at the indicated times after cycloheximide treatment, and levels of UL94 and UL99 were analyzed by Western blotting. (D) 293T cells were transfected with plasmid DNA expressing HAUL94 alone or cotransfected with FLAGUL99, as indicated. Cells were treated with cycloheximide and MG132 at 48 h posttransfection, as indicated. Total protein was harvested at the indicated times after treatment, and levels of UL94 and UL99 were analyzed by Western blotting.