Sanguinarine Modulates Platelet Ca2+ Flux

Intracellular Ca²⁺ flux was measured by flow cytometry [19 (link)]. Washed human platelets (5 × 10⁷/mL) were labeled with 1 μM Fluo-3AM at 37 °C for 30 min, then incubated with sanguinarine (1, 2.5 and 5 μM) or DMSO (control) for 10 min. After collecting a baseline reading for 20 s, collagen (2 µg/mL) and 2 mM extracellular calcium were added to the FACS tube, and the change in fluorescence intensity was recorded on a BD Biosciences flow cytometer (BD Biosciences, Franklin Lakes, NJ, USA) and analyzed using Flowjo V10 (BD Biosciences, Franklin Lakes, NJ, USA). The fitting curve was plotted using SigmaPlot software 14.0 (Systat Software, Inc., San Jose, CA, USA).