PFI-3 (E)-1-(2-hydroxyphenyl)-3-((1R,4R)-2-pyridin-2-yl-2,5-diazabicyclo[2.2.1] heptan-5-yl) prop-2-en-1-one, ODQ 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one, thapsigargin (TG) and phenylephrine (PE) were purchased from Sigma-Aldrich (Saint Louis, MO, USA). Acetylcholine chloride (Ach) was purchased from Harvest Pharmaceutical Co. Ltd. (Shanghai, China). L-NAME, N(ω)-nitro-L-arginine methyl ester, glibenclamide (Gli), and tetraethylammonium (TEA) were purchased from MedChemExpress LLC (Shanghai, China). Fluo-3/AM was purchased from Life Technologies (Invitrogen, Waltham, MA, USA). Ach, PE, physiological salt solution (PSS), and high-K+ salt solution (KPSS) were dissolved in double distilled water, and PFI-3, ODQ, L-NAME, TG, Gli and TEA were dissolved in DMSO (Tianjin Fuyu Fine Chemical Co., Ltd, Wuqing District, China). Arterial smooth muscle cells (A10) were purchased from the American Type Culture Collection (ATCC). KPSS, was composed of (in mM): 74.4 NaCl, 60 KCl, 1.17 MgSO4 ⋅ 7H2O, 1.18 KH2PO4, 14.9 NaHCO3, 1.6 CaCl2, 5.5 D-glucose, and 0.026 EDTA. The PSS solution was composed of (in mM): 130 NaCl, 4.7 KCl, 1.17 MgSO4 ⋅7H2O, 1.18 KH2PO4, 14.9 NaHCO3, 1.6 CaCl2, and 5.5 D-glucose.
L name
Manufactured by MedChemExpress
Sourced in United States
L-NAME is a laboratory reagent used in research settings. It is a nitric oxide synthase inhibitor that can be utilized in experiments to investigate the role of nitric oxide in various biological processes. The core function of L-NAME is to block the production of nitric oxide, but its specific applications are dependent on the research context and objectives.
Automatically generated - may contain errors
Lab products found in correlation
Thapsigargin tg, by Merck Group (1 mentions)
1h 1 2 4 oxadiazolo 4 3 a quinoxalin 1 one odq, by Merck Group (1 mentions)
Fluo 3 am, by Thermo Fisher Scientific (1 mentions)
Phenylephrine pe, by Merck Group (1 mentions)
Verapamil, by MedChemExpress (1 mentions)
Indomethacin, by MedChemExpress (1 mentions)
Glibenclamide, by MedChemExpress (1 mentions)
Ly294002, by MedChemExpress (1 mentions)
Y 27632, by MedChemExpress (1 mentions)
L name, by Merck Group (1 mentions)
4 protocols using l name
1
Vascular Smooth Muscle Cell Calcium Regulation
2
Probing Vasodilation Mechanisms
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To elucidate the role of the endothelium, K+ channel, and PI3K/Akt/Rho-kinase pathways in GPS-mediated vasodilation, the aortic rings with intact endothelium were preincubated with nitric oxide synthase (NOS) inhibitor (100 μM L-NAME), cyclooxygenase (COX) inhibitor (100 μM indomethacin), and soluble guanylyl cyclase (sGC) inhibitor (100 μM methylene blue), respectively, for 30 min, while the aortic rings without endothelium were preincubated with different K+ channel blockers of tetraethylammonium chloride (TEA, 10 mM), 4-aminopyridine (4-AP, 1 mM), BaCl2 (1 mM), glibenclamide (0.01 mM), L-type Ca2+ channel inhibitor (100 μM verapamil), PI3K/Akt inhibitor (10 μM LY294002), and Rho-kinase inhibitor (10 μM Y27632) for 30 min (all inhibitors were obtained from MedChem Express, NJ, USA). After achieving a plateau of PE-induced contracted tension, GPS was added in a cumulative manner (20, 80, and 160 μΜ) for 20 min, and the concentration-response curves were recorded.
3
Coculture of VSMCs and ECs for Nitric Oxide Signaling
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For coculture of VSMCs and ECs, VSMCs were plated on the bottom chamber of transwell cell culture system (pore size 0.4 μm; CoStar Corp) using DMEM with 10% FBS. The next day, the VSMCs were washed and incubated in serum-free media overnight. ECs were cultured on the top chamber of the transwell and allowed to grow until confluence and then placed into the 12-well or 96-well plate of which bottom contained VSMC to initiate the experiments. ECs were transduced with lentivirus to knockdown or overexpress JMJD3 or lentivirus packaging with empty vector used as control. After 48 h, ECs were cultured in fresh EC medium with 2% FBS in addition of 100 μM SNP (MedChemExpress; catalog no.: 14402-89-2) or 300 μM L-NAME (MedChemExpress; catalog no.: HY-18729A) for another 48 h.
4
Induction of HFpEF in C57BL/6J Mice
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Eight-week-old male C57BL/6J mice were used for the establishment of the HFpEF model. The mice were housed in a standard environment with a 12-h light/dark cycle and free access to food and water. To induce HFpEF, a 60% HFD, combined with 0.5 g/l L-NAME (MilliporeSigma, USA) administered through the drinking water, were used. To investigate the role of short-term iNOS inhibition, L-NIL (MedChemExpress, USA) was administered intraperitoneally at a dose of 80 mg/kg of body weight for 3 days after 8 weeks of HFD and L-NAME, as previously described [7] (link). iNOS-null (iNOS-/-) mice were used to examine the long-term effects of iNOS inhibition on HFpEF. All experimental procedures were approved by The Chongqing Medical University Committee (Number of permit:SYXK 2018–0003) on Animal Care and conformed to the Guide for the Care and Use of Laboratory Animals published by the National Institutes of Health.
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