Effect of MG on platelet adhesion, spreading and <i>in vivo</i> thrombus formation.

<p>(a) Representative pictures and (b) quantification of adherent and spread washed human platelets (to fibronectin (Fn)- or collagen (coll)-coated slides) pre-treated with either solvent (CTL) or methylglyoxal (MG, 1 mmol/L, 15 minutes). (c) Representative pictures (upper panel) and quantification (lower graphs) of the effect of <i>in </i><i>vivo</i> treatment of healthy mice with MG (1 mmol/L, 15 minutes) on thrombus size and time to peak after FeCl₃-induced injury of carotid artery. The graphs summarize data obtained in platelets from 12 subjects or 6 animals per group; *P<0.05, ***P<0.001, versus CTL.</p

Effect of MG on platelet [Ca²⁺]_i and degranulation.

<p>(a) Increase in [Ca²⁺]_i measured in washed human platelets treated with either solvent (CTL) or methylglyoxal (MG, 1 mmol/L, 15 minutes) prior to the stimulation with thrombin. (b) Effect of MG pre-treatment on the thrombin (0.03U/ml)-induced release of ATP and (c) on the TRAP-induced surface expression of P-selectin. The graphs summarise the data from at least 6 different individuals; *P<0.05, **P<0.01 versus CTL.</p

Effect of MG on the phosphorylation of β3 integrin and Akt.

<p>(a) Effect of MG (MG, 1 mmol/L, 15 minutes) on fibronectin (Fn) and collagen (coll)-induced tyrosine phosphorylation of β3 integrin (Tyr747). (b) Effect of MG on thrombin -induced tyrosine phosphorylation (Tyr747) of β3 integrin in washed human platelets. (c) Effect of MG on fibronectin (Fn) and collagen (coll)-induced phosphorylation of Akt (Ser 473). (d) Effect of wortmannin (Wt, 20 nmol/L, 30 minutes) on fibronectin (Fn) and collagen (coll)-induced phosphorylation of β3 integrin (Tyr747) and Akt (Ser 473). The graphs summarise the data from 6 different experiments; *P<0.05, ***P<0.001 versus sol or CTL and ^# P<0.05, ^{# # #} P<0.001 versus agonists.</p

Effect of MG on PKC activation.

<p>(a) Membrane translocation of PKCα and β in washed human platelets stimulated with either methylglyoxal (MG, 1 mmol/L, 15 minutes) or thrombin (0.03U/ml) alone or in combination. (b) Effect of methylglyoxal (MG, 1 mmol/L, 15 and 30 minutes) on the phosphorylation of MLC20 in the absence or in the presence of the PKC inhibitor Ro-318820 (Ro, 300 nM, 30 minutes). (c) Effect of MG on thrombin-induced phosphorylation of MLC20. (d) Effect of Ro-318220 on the thrombin-induced aggregation of washed human platelets treated or not with MG. The graphs summarise the data from 6-8 different experiments; *P<0.05, **P<0.01 versus CTL and ^# P<0.05, ^{# #} P<0.01 versus agonists.</p