Figure 3.

Contribution of G_q signaling to Akt translocation in human and murine platelets. (A) Washed human platelets were stimulated with AYPGKF (500 μM) in the presence or absence of 50 nM YM254890, a G_q inhibitor. The effect on Akt translocation and phosphorylation (Ser473 and Thr308) was evaluated by using western blot analysis of platelet membrane fractions using anti-Akt mouse mAb (Santa Cruz Biotechnologies). β₃-integrin was used as the lane loading control. (B) Washed wild-type (WT) and Gα_q-deficient (knockout [KO]) murine platelets were stimulated at 37°C for 2 minutes with AYPGKF (500 μM). Equal amounts of proteins from membrane fractions were analyzed for Akt translocation by western blot analysis using anti-Akt rabbit polyAb (Cell Signaling Technology). (C) Washed human platelets were stimulated with 2MeSADP (100 nM) in the presence or absence of 50 nM YM254890. The effect on Akt translocation and phosphorylation (Ser473 and Thr308) was evaluated by using western blot analysis of platelet membrane fractions using anti-Akt mouse mAb (Santa Cruz Biotechnologies) in panels A and C, and anti-Akt rabbit polyAb (Cell Signaling Technology) in panel B. The western blot analysis shown is representative of 3 independent experiments.