Effect of inhibiting glycolysis on thrombin stimulated platelets.

<p>Platelets were plated on Cell-Tak coated XF96 plates, and pre-treated with 2DG (120 mM) for 1h before the bioenergetic assay. (A) Basal OCR of platelets were measured prior to thrombin injection (0.5 U/ml), followed by 1 μg/ml oligomycin (O), 0.6 μM FCCP (F) and 10 μM antimycin A (AA). (B) Indices of mitochondrial function, basal, thrombin responsive, ATP-linked, proton leak, maximal, reserve capacity and non-mitochondrial OCR were calculated. (C) Basal and thrombin responsive ECAR were calculated from parallel ECAR measurements. Platelets were pre-treated with koningic acid (10 μM) for 1h prior to the bioenergetic assay. (E) Change in OCR after thrombin injection presented as a percentage of control. Data expressed as mean±SEM from one representative one donor, n = 3–5 replicates per sample. *p<0.01, different from control. %p<0.05, different from control. #p<0.01, different from thrombin. &p<0.05, different from thrombin. $p<0.01 different from 2DG.