FIG 6.

The amino terminus of γ₁34.5 is required to form a complex with PKCδ and p32. (A) HeLa cells were infected with the indicated viruses at 5 PFU per cell. At 16 h postinfection, cells were harvested and subjected to immunoprecipitation (IP) with anti-γ₁34.5 antibody. Precipitated proteins and whole-cell lysates (WCL) were processed for Western blot analysis with antibodies against PKCδ, p32, γ₁34.5, and tubulin. Tubulin was used as a loading control. (B) HeLa cells were infected as for panel A and processed for immunoprecipitation with anti-p32 antibody, which was followed by Western blotting with antibodies against PKCδ, p32, γ₁34.5, and tubulin. (C) HeLa cells were transfected with a vector plasmid or a plasmid expressing wild-type γ₁34.5 or the γ₁34.5 mutant devoid of amino acids 1 to 146 (DN) or amino acids 147 to 263 (DC). At 36 h after transfection, cells were processed for immunoprecipitation with anti-γ₁34.5 antibody. Precipitated proteins and whole-cell lysates were probed with antibodies against p32, PKCδ, γ₁34.5, and tubulin.