Fig 8. Aggregation of platelets stimulated with thrombin in the presence of metabolic inhibitors.

Platelets (40 x 10⁶) in suspension were plated onto 96 well plate reader plates, and change in light transmittance following thrombin (0.5 U/ml) addition was measured in platelets subjected to either 1h or 3h treatments consistent with bioenergetic assays, under the following conditions (A) antimycin A (AA, 10 μM), 2DG (120 mM), koningic acid (KA, 10 μM), AA+2DG together and KA+2DG together (B) presence and absence of Gln in media, etomoxir (25 μM) alone and in combination and TMZ (500 μM) and Aza (25 μM) alone and in combination (C) depletion of Gln, etomoxir, TMZ, and Aza together with 2DG. Extent of aggregation is expressed as a percentage of thrombin 0.5 U/ml. Data expressed as mean±SEM from one representative donor, n = 3 replicates per sample. *p<0.01, different from thrombin. #p<0.01, different from thrombin + AA. $p<0.01, different from thrombin + 2DG.