Effect of CGS15943 on CGS21680-induced inhibition of fMLP-stimulated translocation of PKC, RhoA, and ARF1 to membranes.

Unlabeled neutrophils were prewarmed 5 minutes at 37°C. Then 10 μmol/L CB was added and the cell suspensions incubated for an additional 5 minutes in the absence of ADA. Where indicated, 0.1 μmol/L CGS21680 (lanes 3-8) and increasing concentrations of CGS15943 (lanes 4-8) were added to the cell suspensions. Neutrophils were stimulated with 0.1 μmol/L fMLP for 2 minutes (lanes 2-8). Incubations were stopped and neutrophil membranes were prepared as described in “Materials and Methods.” The levels of PKCα, RhoA, and ARF1 in samples were analyzed by immunoblotting. The data are from 1 experiment representative of 3 similar experiments.