FIG. 6.

Coimmunoprecipitation of VP16, VP22, or UL11 with TAP/gE fusion proteins. HaCaT cells were coinfected with Ad vectors expressing TAP/gE550, TAP/gE519, TAP/gE495, TAP/gE470, TAP scrambled, and AdTet trans or were not infected with an Ad vector (No TAP) for 18 h. The cells were subsequently infected with HSV F-gEΔCT and harvested 12 h later in 0.5% NP-40 lysis buffer. Cell extracts were incubated with IgG-Sepharose and washed, and proteins were eluted and subjected to electrophoresis before transfer to PVDF membranes. Membranes were probed with rabbit polyclonal antibodies specific for VP16 (A), VP22 (B), UL11 (C), or anti-protein A antibodies to detect TAP proteins (D). Note that IgG eluted from IgG-Sepharose and the heavy chain comigrated with TAP proteins, as indicated in panel D. VP22 and UL11 levels were quantified using IP Lab Gel software and compared to immunoprecipitation levels for extracts containing no TAP proteins (No TAP), which were set at 1.