U46619 induces transient activation of PKC.
Platelets were labeled with 32P-orthophosphate for 1 hour prior to stimulation with 12 nM convulxin (as a control) and 3 μM U41669. (A) At various time intervals samples were removed to determine the degree of pleckstrin phosphorylation, as described in “Materials and methods.” Results are shown as the mean ± SEM of 3 independent experiments. (B) Washed platelets were stimulated with PMA (30 nM), convulxin (12 nM), or thrombin (0.1 U/mL) for 10 seconds in the presence of indomethacin and apyrase. A sample was removed and platelets were then treated with Ro 31-8220 (1 μM), with further samples being removed for up to 5 minutes (as indicated). After lysis, platelet whole cell lysates were analyzed by Western blotting (not shown) using the pS916 antiserum, and results were analyzed using densitometry to observe active PKD. Results are expressed as the mean ± SEM of 3 separate experiments.
