Figure 8.
SR12813 inhibits tyrosine phosphorylation of SFKs proximal to GPVI, CLEC-2 and integrin αIIbβ3 receptors. Platelets (4 × 108 cells/ml) were pre-treated with vehicle-control (DMSO 0.1% v/v) or SR12813 (0, 50 and 100 μM) for 20 minutes and stimulated for (A, B) 90 seconds with CRP-XL (1 μg/ml) or (C) 120 seconds with rhodocytin (100 nM) in the presence of indomethacin (20 μM), cangrelor (1 μM), MRS2179 (100 μM) and EGTA (1 mM). (D) Washed platelets (4 × 108 cells/ml), pre-treated with SR12813 (0, 50 and 100 μM) or vehicle-control were exposed to fibrinogen-coated wells (100 μg/ml) of a tissue culture plate and allowed to adhere for 30 minutes. Samples were tested for Src (Y418) or Lyn (Y396) phosphorylation. Representative immunoblots are shown. The phosphorylation levels were quantified and expressed as a percentage of untreated (vehicle) controls. Actin was used as a loading control. Full length blots are shown in supplementary Fig. 9 and 10. Results are mean ± SD (n ≥ 3), *P < 0.05, **P < 0.01, ***P < 0.001 and ****P < 0.0001 was calculated by one-way ANOVA. Figure adapted from corresponding PhD thesis48.