Figure 4.

Interaction of PhLP with Gβγ but not with CCT is necessary for Gβγ dimer assembly. (A) HEK-293 cells were transiently transfected with PhLP-myc, PhLP 132–135A, PhLP Δ1–75, PhLP/Pdc 76–94, or empty vector as indicated along with Gβ₁ and HA-Gγ₂ and then assayed after 48 h. The binding of PhLP variants to Gβγ and CCT was measured by immunoprecipitating with an antibody to the C-terminal myc tag and immunoblotting for PhLP, Gβ or CCTɛ. Blots are representative of similar data from three experiments. (B) Levels of overexpressed Gβ_lγ₂ dimer were determined by immunoprecipitation as in Figure 2A. Bars represent the average±s.e. from three separate experiments (^*P<0.01, ^**P<0.001). (C) The rate of nascent Gβ_lγ₂ dimer formation was determined by a pulse–chase assay as in Figure 2B. Symbols represent the average±s.e. from three separate experiments.