Fig. 5

BML-111 attenuates thrombosis. A lipophilic dye, DiOC6 (5 µM) was added to citrated human blood and incubated for 1 h at 30 ℃, then treated with BML-111 (25 and 50 µM) or a vehicle-control (modified-Tyrode’s HEPES buffer) for 5 min. The labelled blood was then perfused across Vena8 Biochips, coated with collagen (100 µg/ml) under arterial flow conditions (shear stress: 20 dynes/cm²). (a) Representative images show thrombus formation over 540 s in 25 and 50 µM BMLL-111 or vehicle-control treated samples. Fluorescence was measured using an excitation wavelength of 488 nm and emission at 500–520 nm with an argon laser. A Nikon A1R confocal microscope (20x objective) was used to observe thrombus formation, and images were captured every second for over 600 s. (b) Quantified data represents mean thrombus fluorescence intensity for BML-111 and vehicle-control treated samples. The data were analysed using NIS-Elements software and normalised to the fluorescence level at the end of the assay in the vehicle-control treated sample. Data represent the mean ± SEM (n = 4), ****P ≤ 0.0001 was as calculated by Two-way ANOVA.