Experimental validation of computerised models of clustering of platelet glycoprotein receptors that signal via tandem SH2 domain proteins
Fig 9
Synergistic inhibition of platelet aggregation by CLEC-2 and GPVI agonists by threshold concentrations of blocking biologics and the Syk inhibitor PRT-060318.
(Ai) Representative traces of washed platelets (2x108 platelets/ml) preincubated for 10 min with threshold concentrations of PRT-060318 (2.5 nM) and/or the divalent anti-CLEC-2 antibody fragment, AYP1 F(ab)2 (2.2 nM). The arrow indicates the time of addition of rhodocytin (100nM). (Aii) The mean ± s.d. level of aggregation (%) after 10 min (n = 3). (Bi) Representative traces of washed platelets (2x108 platelets/ml) preincubated for 10 min with non-inhibitory concentrations of PRT-060318 (2.5 nM) and/or the divalent anti-GPVI nanobody, Nb2-2 (2.2 nM). The arrow indicates the time of addition of CRP (1 μg/ml). (Bii) The mean ± s.d. level of aggregation (%) after 3 min (n = 3: in one donor, 0.50 nM Nb2-2 was used due to 2.2 nM causing complete inhibition). Significance was measured using one-way ANOVA with a Tukey post-hoc test where P ≤ 0.05.