Effect of Gln on thrombin stimulated platelets.

<p>Platelets were plated on XF96 plates Cell-Tak coated, and incubated in either regular XF DMEM media or media without Gln for 1h, before bioenergetic measurements. (A) Basal OCR of platelets were measured ahead of 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. Bioenergetic assays were performed in XF DMEM media containing different concentrations of Gln (0–4 mM). (D) Basal OCR prior to thrombin injection, thrombin linked, ATP linked and maximal OCR after thrombin injection presented as a percentage of highest concentration of Gln (4 mM) after subtraction of no Gln OCR. Platelets were pre-treated with Aza (0–50 μM) for 3h before 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<0.01, different from thrombin. $p<0.01 different from Ctrl-no glut. @p<0.01, thrombin linked OCR different from Gln (500 μM). %p<0.01, ATP-linked OCR different from Gln (500 μM). +p<0.01, maximal OCR different from Gln (500 μM).