![Figure 4. Effect of PI3K activity/Gi pathway on Akt translocation in human and murine platelets. Washed human platelets were stimulated with AYPGKF (500 μM) (A), SFLLRN (5 μM) (B), or 2MeSADP (100 nM) (C) in the absence or presence of LY294002 (25 μM; a pan-PI3K inhibitor). (D) Washed human platelets were stimulated with AYPGKF (500 μM) in presence or absence of ARC69931MX (100 nM; a P2Y12 antagonist). The effects of inhibitor/antagonist on Akt translocation and phosphorylation were determined by western blot analysis. The data shown are representative of 3 experiments. (E) WT and P2Y12-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. The western blot analysis shown is representative of 3 independent experiments using anti-Akt rabbit polyAb (Cell Signaling Technology) in panels A and E, and anti-Akt mouse mAb (Santa Cruz Biotechnologies) in panels B-D.](https://ash.silverchair-cdn.com/ash/content_public/journal/blood/125/1/10.1182_blood-2014-05-576306/4/175f4.jpeg?Expires=1735817588&Signature=D2YxLYLPERMWN4nUa4J1T3KmEodU6WAajMqrLt7hEe2i3ObqlHVU0rXDE-RIyCaqdXTUethdc9G9JsRC9rdnjOR2VxUI0X8EX~lsCNK1nzACt-jJbrN9lYCE6fJmK~BEgFZ3CZ-TXCIedvcH4aYAUvnK6UVmu7uJArEDteC-i3Lj~11ZCdVhwJdekZAbZFhNIu5HIpUxSghElamqw63prrCa0vIqNvVjuQUekoL3YqjTSwezYzU4SWB6Z8RKrPJA-F1YkyEaraPzkeihJyQ4NaDjpxYzN2SKFOimk~LKI51sp67VrcDGKZoDRszVNVXbIbjAEmDWoNftRHx7Ft3VBQ__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
Effect of PI3K activity/Gi pathway on Akt translocation in human and murine platelets. Washed human platelets were stimulated with AYPGKF (500 μM) (A), SFLLRN (5 μM) (B), or 2MeSADP (100 nM) (C) in the absence or presence of LY294002 (25 μM; a pan-PI3K inhibitor). (D) Washed human platelets were stimulated with AYPGKF (500 μM) in presence or absence of ARC69931MX (100 nM; a P2Y12 antagonist). The effects of inhibitor/antagonist on Akt translocation and phosphorylation were determined by western blot analysis. The data shown are representative of 3 experiments. (E) WT and P2Y12-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. The western blot analysis shown is representative of 3 independent experiments using anti-Akt rabbit polyAb (Cell Signaling Technology) in panels A and E, and anti-Akt mouse mAb (Santa Cruz Biotechnologies) in panels B-D.
