Figure 6
SR12813 inhibit outside-in signalling in platelets. Human washed platelets (2 × 107 cells/ml) were treated with SR12813 (50 and 100 μM) or vehicle-control (DMSO 0.1% v/v) for 20 minutes and added onto (A) fibrinogen (100 μg/ml) or (C) collagen-coated coverslips for 45 mins. Platelets were stained using Alexa-Fluor 488 for visualisation using a Nikon A1-R confocal microscope (100X). 5 images were captured of each sample at random locations. Representative images of platelet adhesion and spreading are shown. Cumulative data of platelets adhered in each sample is shown. Spreading platelets were divided into 3 classes: (adhered but not spread; filopodia: spreading platelets and lamellipodia: fully spread). Results expressed (as relative frequency) as the percentage of the total number of platelets adhered. (B) Human PRP was incubated with SR12813 (20, 50 and 100 μM) or vehicle-control (DMSO 0.1% v/v) for 20 minutes. Extent of clot retraction was determined by comparing clot weight after 60 minutes. Representative image of clot retraction after the end of the assay is shown. Cumulative data represent clot weight (in mg) of samples treated with SR12813 compared with vehicle-control. Data represent mean ± SD (n ≥ 3), *P < 0.05, **P < 0.01 and ***P < 0.001 was calculated by one-way ANOVA. Figure adapted from corresponding PhD thesis48.
