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. Author manuscript; available in PMC: 2019 Jun 1.
Published in final edited form as: J Thromb Haemost. 2018 May 8;16(6):1211–1225. doi: 10.1111/jth.14005

Figure 5. Genetic ablation of PDK1 reduces 2MeSADP induced platelet aggregation, and abolished MAPK pathway mediated thromboxane generation.

Figure 5

(A) Washed platelets from pdk1fl/fl and pdk1−/− mice were stimulated with 2MeSADP (50 nM) under stirred conditions at 37°C for 5 min. Aggregation was measured using light transmission aggregometry. Light transmission and area under the curve (mean ± SEM) are calculated from six independent experiments. (B) Washed platelets from pdk1fl/fl and pdk1−/− mice were stimulated with 2MeSADP (50 nM) under stirred conditions for 5 min at 37°C. Thromboxane generation was measured using ELISA. Graphs represent mean ± SD from at least three different experiments (P=0.0014). (C) Washed platelets from pdk1fl/fl and pdk1−/− mice were stimulated with 2MeSADP (50 nM) under stirred conditions for 3 minutes. Platelet proteins were separated by SDS-PAGE, immunoblotted and probed for phospho Raf1 (Ser338), MEK1/2 (Ser217/221), ERK1/2 (Thr202/Tyr204), cPLA2 (Ser505) and Akt (Thr308). The Western blot shown is a representative of three independent experiments.