2.6 Materials
Physiological salt solution (PSS) contained (in mM) NaCl (115), NaHCO3 (25), K2HPO4(2.5), MgSO4 (1.2), Glucose (5.5), HEPES (10) and CaCl2 (1.3). For PSS containing 32 instead of 5 mM K+, the NaCl concentration was reduced to 88 mM and 27 mM KCl was added. Buffers where continuously aerated with 5% CO2 in air at 37 °C. Bradykinin (BK; Sigma-Aldrich) was dissolved in 0.1 M acetic acid. 2-(4-Carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide potassium salt (c-PTIO; Sigma-Aldrich) and 7-nitroindazole (7-NI; Sigma-Aldrich), 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ; Cayman Chemical, Ann Arbor, Mi, USA) and N-hydroxy-2-(methylsulfonyl)benzenesulfonamide (CXL-1020; Axon Medchem, Groningen, NL) were dissolved in DMSO. Indomethacin (INDO; Sigma-Aldrich) and N-omega-propyl-L-arginine (NPLA; Sigma-Aldrich) were dissolved in 100% EtOH. CXL-1020 (Axon Medchem, Groningen, The Netherlands), endothelin-1 (ET-1; Bachem, Weil am Rhein, Germany), hydrogen peroxide (H2O2; Sigma-Aldrich), N-omega-nitro-L-arginine methyl ester (L-NAME; Sigma-Aldrich) and sodium nitroprusside (SNP; Sigma-Aldrich) were dissolved in ddH2O. 3-Amino-1,2,4-triazole (Amitrole; Sigma-Aldrich), catalase (CAT; Sigma-Aldrich), 3,17-diacetoxyestra-1,3,5(10)-trien-2-carboxylic acid (DETCA; Sigma-Aldrich) and N-acetylcysteine (NAC; Sigma-Aldrich) were dissolved in PSS. The biological effects and concentrations of the pharmacological tools that we used are summarized in Table 2.