4 hydroxycoumarin

Coumarin and 4-Hydroxycoumarin were obtained from Acros organics, 3-Hydroxycoumarin, 3-(2-Hydroxyphenyl) propionic acid, and 7-hydroxycoumarin (7OHC) were obtained from Sigma-Aldrich, trans-o-Coumaric acid was obtained from Tokyo chemical Industry Co., and 6-Hydroxycoumarin was obtained from Fisher scientific. All chemicals had purities of 98 % or higher. Methanol (≥99.8 %) and acetic acid (≥99.8 %) were procured from Fisher Scientific and Sigma Aldrich, respectively. All chemicals were used as received. Stock solutions of coumarin (8.2 mM) and its products (100 μM) were prepared to their desired concentration and diluted using distilled water when required. Calibration curves for each chemical were prepared using their associated stock solution and analysed using high performance liquid chromatography (HPLC). HPLC mobile phases were prepared using deionised water. The initial concentration of coumarin for this study was 100 µM (15 ppm), as followed in our previous study pertaining to dosimetry [26] .

Chemicals are commercially available: DCPIB (Sigma-Aldrich, #SML2692-25MG), ARL 67156 trisodium salt (Tocris, #1283), S1P (Cayman, #62570), W146 (Tocris, #36024), JTE013 (Tocris, #2392), BAPTA AM (Cayman, #15551), diphenyleneiodonium (Tocris, #0504), ATP (Sigma-Aldrich, #A2382-5G), lipopolysaccharide (LPS) (Sigma-Aldrich, #L2880-100MG), nigericin (Cayman, 11437), dicumarol (Acros Organics, #204120050), chenodiol (VWR 21947-5G), tioxolone (Sigma-Aldrich, #217077), tolfenamic acid (Sigma-Aldrich, #T0535-5G), flufenamic acid (Acros Organics, #165920100), zafirlukast (TCI America, #Z0029-100MG), 4-hydroxycoumarin (Acros Organics, #121100050), and warfarin (Agilent, #PST-1000). All chemicals were dissolved with the manufacturers’ instructions. Dicumarol stock solution (10 to 20 mM) was made in dimethyl sulfoxide with brief sonication (2 to 5 min) and heating (50°C).