Cells were treated with 10 μM rabeprazole (Rabe) or omeprazole (Ome) to inhibit PPs (Selleck, Shanghai, China). Cells were treated with 10 μM Cytochalasin D to inhibit phagocytosis (MedChemExpress, Shanghai, China). Cells were treated with 10 μM indomethacin (MedChemExpress) or chlorpromazine (Selleck) to inhibit caveolae‐ or clathrin‐mediated endocytosis, respectively. Macropinocytosis was inhibited by treatment cells with 10 μM LY294002 or amiloride (Selleck). Cells were treated with 10 μM KM91104 or 10 nM Baf‐A1, or 10 μM azathioprine (Selleck) to inhibit the activity of v‐ATPase or RAC1, respectively. Cells were treated with 10 μM Dynasore (MedChemExpress), CDC42‐IN‐1 (Selleck) or NAV‐2729 (MedChemExpress) to inhibit the activity of dynamin, CDC42 and ARF6, respectively. Cell cholesterol was extracted by treatment with 10 μM MeβCD (Aladdin). Equal pH gradients in cells were induced by 10 μM 2, 4‐DNP (Merck). Cells were treated with 10 μM H89 (Selleck) to inhibit PKA activation. In most experiments, cells were pretreated with the above reagents for 2 h, followed by subsequent experiments.
Indomethacin
Manufactured by MedChemExpress
Sourced in United States
Indomethacin is a nonsteroidal anti-inflammatory drug (NSAID) used as a laboratory reagent. It functions as a cyclooxygenase (COX) inhibitor, which can help in the study of inflammatory pathways and processes.
Automatically generated - may contain errors
Lab products found in correlation
Bafa1, by Selleck Chemicals (1 mentions)
Rabeprazole, by Selleck Chemicals (1 mentions)
Amiloride, by Selleck Chemicals (1 mentions)
Omeprazole, by Selleck Chemicals (1 mentions)
Chlorpromazine, by Selleck Chemicals (1 mentions)
Azathioprine, by Selleck Chemicals (1 mentions)
Cytochalasin d, by MedChemExpress (1 mentions)
Dynasore, by MedChemExpress (1 mentions)
Ly294002, by Selleck Chemicals (1 mentions)
Verapamil, by MedChemExpress (1 mentions)
L name, by MedChemExpress (1 mentions)
Glibenclamide, by MedChemExpress (1 mentions)
Ly294002, by MedChemExpress (1 mentions)
Y 27632, by MedChemExpress (1 mentions)
B27 supplement, by Thermo Fisher Scientific (1 mentions)
Ham s f 12 nutrient mix, by Thermo Fisher Scientific (1 mentions)
Recombinant human il 1β, by R&D Systems (1 mentions)
Glutamine, by Thermo Fisher Scientific (1 mentions)
Neurobasal a, by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin solution, by Thermo Fisher Scientific (1 mentions)
6 protocols using indomethacin
1
Inhibition of Endocytic Pathways in Cells
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
Neuron-Mesenchymal Stem Cell Co-Culture
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For each individual experiment, a minimum of 20 mice were anesthetized and performed transcardial perfusion with cold standard, bubbled with 95% O2–5% CO2, artificial cerebrospinal fluid (ACSF) solution.50 (link) Neurons from vagus ganglia could be collected and placed in ice-cold Ca2+ and Mg2+-free Hank’s Balanced Salt Solution supplemented with penicillin–streptomycin. The culture medium consisted of 21.5 mls of Ham’s F-12 Nutrient Mix (GIBCO); 21.5 mls of Neurobasal A (GIBCO); 5mls Glutamine; 1 ml of B-27 Supplement (GIBCO); 1 ml of Penicillin-Streptomycin solution (Thermo). The ratio of MSCs to neurons was 10:1. Recombinant human IL-1β (100 ng/ml, R&D systems) and mouse IL-1β (100 ng/ml, R&D systems) were used to stimulate the PGE2 production of MSCs. Indomethacin (10 nM, MedChem Express) and MF63 (10 nM, MedChem Express) were used to inhibit PGE2 synthetase, PTGS2 and PTGES, respectively.
4
Indomethacin, SAC, and 4-PBA Protocol
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Indomethacin (99.71%) was obtained from MedChemExpress LLC (Monmouth Junction, NJ, USA). SAC (≥ 98%) and 4‐phenyl butyric acid (4‐PBA; 99%) were obtained from Sigma‐Aldrich (St. Louis, MO, USA). Indomethacin was dissolved in DMSO, subpacked, and stored at −80 °C. SAC and 4‐PBA were dissolved in PBS, respectively, and aliquots were stored at −20 °C.
5
Lipofectamine 2000-Mediated siRNA Transfection
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The Lipofectamine 2000 reagent was obtained from Invitrogen (Life Technologies Corporation, Carlsbad, CA). The primers for Cyp2C9 siRNA, and their controls were purchased from RiboBio (Guangzhou, China). The PPARα ligand AVE8134, 2-Methyl-6-({3-[(2-phenyl-1,3-oxazol-4-yl)methoxy]propoxy}methyl) benzoic acid, were synthesized by Dr. John R. Falck and kindly offered by Jorge H. Capdevila from the Department of Medicine (Division of Nephrology), Vanderbilt University, Nashville, USA. Wyeth-14,643, Bezafibrate, the PPARα antagonist GW6471, and the COX inhibitor indomethacin were purchased from MedChemExpress (New Jersey, USA). 11-HETE and four kinds of EETs were purchased from Cayman Chemical (Ann Arbor, Michigan, USA). For the purchasing information on some of the other conventional reagents in our lab, please refer to our previous articles [15 (link), 20 (link), 21 (link)].
6
Antiviral Drug Screening Protocol
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Acetaminophen (catalog number HY-66005), arbidol hydrochloride (HY-14904A), aspirin (HY-14654), atazanavir sulfate (HY-17367A), baloxavir marboxil (HY-109025), daclatasvir (HY-10466), darunavir (HY-17040), dolutegravir (HY-13238), efavirenz (HY-10572), elbasvir (HY-15789), etravirine (HY-90005), galidesivir (BCX4430, HY-18649A), ganciclovir (HY-13637), glecaprevir (HY-17634), hydroxychloroquine (HY-B1370), indomethacin (HY-14397), ledipasvir (HY-15602), lopinavir (HY-14588), losartan (HY-17512), nafamostat mesylate (HY-B0190A), nitazoxanide (HY-B0217), ombitasvir (HY-13997), oseltamivir (HY-13318), penciclovir (HY-17424), perindopril salt (HY-B0130A), raltegravir (HY-10353), remdesivir (GS-5734, HY-104077), rilpivirine (HY-10574), ritonavir (HY-90001), simeprevir (HY-10241), sofosbuvir (HY-15005), and β-d-N4-hydroxycytidine (HY-125033) were purchased from MedChemExpress (Monmouth Junction, NJ, United States). Amodiaquin dihydrochloride dihydrate (A2799), artesunate (A3731), chloroquine phosphate (PHR1258), mefloquine hydrochloride (PHR1705), peramivir trihydrate (SML2486), ribavirin (R9644), teicoplanin (T0578), and toremifene citrate salt (T7204) were purchased from Sigma-Aldrich (St, Louis, Missouri, United States). Favipiravir (T-705) was provided by the Toyama Chemical Co., Ltd. (Japan). The drug solutions were filtered through a 0.22 μM membrane and were stored at −80°C.
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