AFfirm™ VASP Mouse Monoclonal Antibody - #BF8131

Fig. 6. The effect of FCF on human platelet to agonists stimulation. Washed human platelets were pre-incubated with different doses of FCF (25, 50, and/or 100 µM) at 37 °C for 10 min, subsequently subjected into platelets aggregation, western blotting, adhesion and clot retraction assays in response to stimulation with thrombin (0.01 U/mL), collagen (2 µg/mL) and/or ADP (20 µM). (A) Bars graph showing the quantified inhibition rates of platelet aggregation (n = 3 each group). (B) Representative western blotting showing phosphorylation levels of PI3K, AKT, GSK3β, ERK1/2, p-p38, JNK, VASP (S157) and VASP (S239) (n = 3 each group). (C) Human platelets were subjected on FN matrix for one hour in presence of 0.01 U/mL thrombin, and IF images showing platelet morphological alternation and intracellular CD62P expression (Red, F-actin; Green, CD62P). Bar graphs quantified (D) adherent platelet number, (E) averaged platelet area and (F) CD62P MFI values (n = 3 each group). (G) The representative images showing human platelets colt retract progression and (H) the curve plots summarized relative colt retraction (n = 3 each group). (I) human platelets supernatants from aggregation assay (FCF, 100 µM) were harvested for ATP releases by luciferin/luciferase reagents and the data are presented as the relative percentage to solvent (n = 3 each group). The graphs summarize the data from at least three times independent experiments, data are shown as the mean ± S.E.M., and analyzed using a One-way ANOVA or Unpaired student’s t-test, and *p < 0.05, **p < 0.01, and ***p < 0.001 versus solvent (Sol.). (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)