FIGURE 5.

Co-immunoprecipitation of AKT, ERK1/2, and HA-PLD2 in resting and stimulated cells. HEK293 cells stably expressing fPR were transfected with empty vector (pcDNA3) or plasmid encoding wild-type HA-PLD2. Cells in suspension were treated in the absence or presence of 3 μm AKTib1/2 for 15 min before stimulation with 1 μm fMLP for 60 s. Immunoprecipitation (IP) of AKT (A), ERK1/2 (B), and HA-PLD (C) was performed with cell homogenates, and the presence of partners was revealed by Western blotting. For each immunoprecipitation experiment, the stimulating effect of fMLP was calculated and is expressed as a percentage of stimulated control values (right panels). Data are representative of three independent experiments.