Ucl1684

The following drugs were used: Acetylcholine, phenylephrine hydrochloride (PE), iberiotoxin, SNP (sodium nitroprusside dehydrate), U46619 (9α-epoxymethanoprostaglandin F_2α), pirfenidone (5-methyl-1-phenyl-2-(1H)-pyridone), flupiritine (ethyl 2-amino-6-((p-fluorobenzyl)amino)-3-pyridinecarbamate), mannitol, N^G-nitro-l-arginine (L-NOARG), tempol, TEA (tetraethylammonium), TRAM-34, UCL1684, and XE991 (10,10-bis(4-pyridinylmethyl)-9(10H)-anthracenone) were purchased from Sigma (St Louis, MO, United States). TRAM-34, UCL1684, and XE991 were dissolved in dimethylsulphoxide (DMSO), and further diluted in distilled water. Unless otherwise stated the substances were dissolved in distilled water. The concentration of DMSO in the bath was below 0.01% and to examine whether vehicle affected vascular contractility parallel control curves for the vehicle were obtained.

All drugs and reagents including GSK1016790A, HC-067047, RN-1734, SKA-31, UCL1684 and 4α-PDD were purchased from Sigma (Sigma-Aldrich, Co., Louis, MO, USA). Apamin was purchased from Santa Cruz Biotech (Santa Cruz Biotechnology, Inc., Dallas, Texas, USA). Dimethyl sulfoxide (DMSO) was used to make stock solutions. Final concentration of DMSO in the bath solution was less than 0.01%.

Cremophor EL, dimethyl sulfoxide (DMSO), and urethane were from Sigma-Aldrich (Steinheim, Germany); NaCl was from POCh (Gliwice, Poland). Acetylcholine chloride, (-)- phenylephrine hydrochloride, l-NAME, indomethacin, ouabain, and barium chloride (MP Biomedicals, Santa Ana, CA, USA) were dissolved in deionized water except for indomethacin (dissolved in 0.5 M NaHCO₃). SKA-31 (Abcam, Cambridge, MA, USA) was dissolved immediately before the in vivo experiments in a mixture of DMSO, Cremophor EL and saline (1:2:7) to obtain a concentration of 10 mg/mL and then it was further diluted with saline to obtain final concentrations of 1 and 3 mg/0.5 mL. Stock solutions (10 µM) of UCL1684, NS309 (Sigma-Aldrich, St.Louis, MO, USA), SKA-31 and TRAM-34 (Tocris Bioscience, Bristol, UK) were prepared in DMSO. The final concentrations of these agents were prepared by dilutions with deionized water which adjusted the final concentrations of DMSO to ≤0.1% v/v for experiments on isolated organs. None of the solvents used affected basal parameters in vivo or in vitro.