Ns1619

Experiments were performed on 4–6-month-old spontaneously hypertensive (SHR) rats and normotensive Wistar Kyoto (WKY) rats of either sex. The rats were purchased from Charles River (Wilmington, MA, USA) and housed under controlled conditions with a 12:12 h light–dark cycle. Food and water were available to the animals ad libitum. All animal protocols were approved by the Institutional Animal Care and Use Committee (IACUC) of North Dakota State University.
Paxilline, Angiotensin II (Ang II), and NS1619 were purchased from Cayman (Ann Arbor, ML, USA). Anti-SM actin antibody, Dulbecco’s modified Eagle medium (DMEM), and fetal bovine serum (FBS) were obtained from Thermo Fisher Scientific (Waltham, MA, USA). Collagenase was purchased from Worthington Biochemical (Lakewood, NJ, USA). Anti IP3 receptor (IP3R)1 antibody and protein A/G plus agarose were obtained from Santa Cruz (Dallas, TX, USA). KCNMA1 antibody was purchased from Alomone Lab (Jerusalem, Israel). 4-[3-(4-iodophenyl)-2-(4-nitrophenyl)-2H-5-tetrazolio]-1, 3-benzene disulfonate (WST-1) was obtained from Abcam. 4′,6-diamidino-2-phenylindole (DAPI), Adenophostin A (AdA), Norepinephrine (NE), ATP, GTP, HEPES, and others were purchased from Sigma-Aldrich (St. Louis, MO, USA).

Acetylcholine chloride, sodium nitroprusside, and noradrenaline hydrochloride were obtained from Sigma-Aldrich (St. Louise, MO, USA); potassium chloride from Chempur (Piekary Śląskie, Poland); pinacidil, NS1619, and U-46619 from Cayman Chemical (Ann Arbor, MI, USA). Stock solutions (10 mM) of these drugs were prepared in distilled water, except for noradrenaline, which was dissolved in NaCl (0.9%) + ascorbic acid (0.01% w/v) solution; pinacidil, NS-1619 were dissolved in DMSO, and U-46619 in ethanol.
These solutions were stored at −20 °C, and appropriate dilutions were made in Krebs-Henseleit solution (KH in mmol/L: NaCl 115; CaCl₂ 2.5; KCl 4.6; KH₂PO₄ 1.2; MgSO₄ 1.2; NaHCO₃ 25; glucose 11.1) on the day of the experiment. The maximal solvent concentration in the medium was less than 0.01% (vol/vol). At these concentrations, solvents did not alter the reactivity of the studied arteries.
Hemp seeds were purchased from Ekogram (Zielonki, Poland), and unrefined, cold-pressed hemp seed oil was obtained from Ol’Vita (Panków, Poland).