YM-254890 (henceforth referred to as YM) was purchased from AdipoGen® Life Sciences. The morphine used was sterile and free of preservatives (Morphine Sulphate, Sigma-Aldrich, St. Louis, MO, USA). For all in vivo studies, the vehicle used for YM compound is (0.05% DMSO in 5% dextrose solution) unless 0.9% saline is indicated as for morphine. To observe the systemic effects of YM administration, YM doses (0.1, 0.3, 0.5 and 1 mg/kg) was given subcutaneously. The dose of YM for subcutaneous injection was used from previous published studies (Meleka et al., 2019 (link)). The dose of morphine selected for subcutaneous administration (2.5, 5 and 10 mg/kg) is commonly used in mouse analgesia (Kest et al., 2002 (link)). For the combined administration of morphine and YM on thermal antinociception, YM was administered 10 min before the morphine. The latency of response was measured before injection of drugs (baseline latency response) and at different time intervals (post-treatment latency response) after drugs injection.
Ym 254890
Manufactured by Adipogen
Sourced in United States
YM-254890 is a selective inhibitor of the Gαq protein. It is commonly used in laboratory research to study the Gαq signaling pathway and its involvement in various cellular processes.
Automatically generated - may contain errors
Lab products found in correlation
Firefly d luciferin, by Nanolight Technology (2 mentions)
Endothelin 1 et 1, by Bio-Techne (2 mentions)
Dulbecco s modified eagle s medium dmem, by Thermo Fisher Scientific (2 mentions)
Dulbecco s phosphate buffered saline, by Corning (2 mentions)
Trypsin, by Corning (2 mentions)
Fluorobrite dmem, by Thermo Fisher Scientific (2 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
Bovine serum albumin (bsa), by Merck Group (2 mentions)
Thrombin, by Merck Group (2 mentions)
Enzyme free cell dissociation solution, by Merck Group (2 mentions)
Fetal bovine serum (fbs), by Corning (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Morphine sulphate, by Merck Group (1 mentions)
Clozapine, by Bio-Techne (1 mentions)
Aripiprazole, by Bio-Techne (1 mentions)
Paliperidone, by Bio-Techne (1 mentions)
Risperidone, by Bio-Techne (1 mentions)
Way100635, by Bio-Techne (1 mentions)
Selumetinib azd6244, by Selleck Chemicals (1 mentions)
10 protocols using ym 254890
1
Investigating Thermal Antinociception via YM-254890 and Morphine
2
Pharmacological Reagents for Neuromodulation
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Aripiprazole, clozapine, haloperidol hydrochloride, H89 dihydrochloride, L 741,626, MDL 100907, paliperidone, risperidone, SKF 83822, SCH 39166, TCB-2, WAY 100635, 7-OH-DPAT and 8-OH-DPAT were purchased from Tocris Bioscience (Spain). Forskolin and selumetinib (AZD6244) were purchased from Selleckchem (Spain). Cholera toxin (CTX), pertussis toxin (PTX) and 12-O-tetradecanoylphorbol-13-acetate (TPA) were purchased from Sigma-Aldrich (MO, USA). YM-254890 was purchased from Adipogen Life Sciences (CA, USA). All were dissolved in dimethyl sulfoxide, except CTX and WAY 100635 that were dissolved in water.
3
Cell Culture and Reagent Conditions
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MDA-MB-231 cells were purchased from the American Type Culture Collection (Rockville, MD). NCI-H23, A-498, SNU-423, HeLa, COLO 205, and Calu-3 cells were purchased from the Korean Cell Line Bank (Seoul, Korea). HEK293A cells were purchased from Invitrogen (Carlsbad, CA). MDA-MB-231, NCI-H23, A-498, SNU-423, and COLO 205 cells were cultured in RPMI 1640 supplemented with 10% fetal bovine serum (FBS), 100 U/mL penicillin, and 100 μg/mL streptomycin at 37 °C in 5% CO2. HeLa cells were grown in modified Eagle’s medium (MEM; HyClone, Logan, UT) supplemented with 10% FBS, penicillin, and streptomycin. HEK293A and Calu-3 cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM; HyClone) supplemented with 10% FBS, penicillin, and streptomycin. CXCL12 (#300-28A) was purchased from PeproTech (London, UK). Histamine (#H7250) was purchased from Sigma‒Aldrich (St. Louis, MO). AMD3100 (#HY10046), U73122 (#HY-13419), and U0126 (#HY-12031) were purchased from MedChemExpress (Monmouth Junction, NJ). Pyrilamine (#0660) and PTX (#3097) were purchased from Tocris Bioscience (Ellisville, MO). YM254890 (#AG-CN2-0509-MC05) was purchased from AdipoGen Life Sciences (San Diego, CA).
4
GPR43 Agonist and Antagonist Assay
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The GPR43-selective agonist compound 187 (4-[(2R,6S)-2, 6-dimethylmorpholin-4-yl]-7-(2-fluorobenzenesulfonyl)-2- methyl-5H-pyrrolo[3,2-d]pyrimidin-6-amine), which was invented by Takeda Cambridge Ltd. (https://patentscope.wipo.int/search/en/detail.jsf?docId=WO2015198045 ), was kindly provided by Dr. Y.S. Kwak (Korea University). The selective inverse agonist (S)-3-(2-(3-chlorophenyl)acetamido)-4-(4-(trifluoromethyl)phenyl)butanoic acid (CATPB), sodium acetate, phorbol 12-myristate 13-acetate (PMA), lipopolysaccharide (LPS), and pertussis toxin (PTX) were purchased from Sigma-Aldrich (USA). Cumate solution (10,000×; PBQM100A-1) was acquired from System Biosciences (USA), YM-254890 from Adipogen (USA) and C3 toxin (C3 transferase from Clostridium botulinum, #CT04) from Cytoskeleton (USA).
The GPR43-myc construct was cloned into PiggyBac cumate switch-inducible vector (PB-CuO, PBQM-812-A1) using the restriction sites NheI and NotI (New England BioLabs, USA).
The GPR43-myc construct was cloned into PiggyBac cumate switch-inducible vector (PB-CuO, PBQM-812-A1) using the restriction sites NheI and NotI (New England BioLabs, USA).
5
Regulation of M2 Macrophage Activation by P2Y11 Signaling
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ATPγS (Sigma Aldrich, St. Louis, MO, USA) and NF546 (Tocris, Bristol, UK) were used as P2Y11 receptor agonists (10 to 30 μM). NF340 (Santa Cruz, Dallas, TX, USA) served as P2Y11 receptor antagonist (10 μM) (Gruenbacher et al., 2019). Additional reagents used in this study are as follows: P2X1 receptor antagonist NF449 (10 μM) (Tocris), adenylyl cyclase inhibitor SQ22536 (5 to 20 μM) (Tocris), Gq/11 inhibitor YM‐254890 (0.1 to 0.5 μM) (Adipogen Life Sciences, Liestal, Switzerland), recombinant IL‐1α and IL‐1ß (50 to 1,000 pg·ml−1) (R&D, Bio‐Techne), recombinant IL‐1 receptor antagonist (IRAP ; 0.05 to 0.2 μg·ml−1) (R&D, Bio‐techne); nonselective phosphodiesterase inhibitor 3‐isobutyl‐1‐methylxanthine (IBMX ) (20 to 200 μM) (Sigma‐Aldrich); PDE4‐ selective inhibitor rolipram (2 to 10 μM) (Sigma‐Aldrich), the TACE/ADAM17 inhibitors TAPI‐1 and TAPI‐2 (10 to 50 μM) (Tocris); lipopolysaccharide (LPS) from Salmonella abortus equi (10 pg·ml−1 to 10 ng·ml−1) (Sigma‐Aldrich) was used to activate M2 macrophages via TLR4.
6
Pharmacological Inhibitor Purchasing Protocol
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ACEA, Wortmannin, WIN 55,212-2, and Rapamycin were purchased from Cayman Chemical (USA). PTX was purchased from Sigma-Aldrich (Israel). YM-254890 was purchased from AdipoGen (USA). JD5037 and DO34 were purchased from MedChemExpress (China).
7
CB1/CB2 Receptor Agonist and Antagonist Assays
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RPMI Medium 1640 was from GIBCO/Invitrogen (Grand Island, NY, USA). PCR reagents were from Applied Biosystems. ACEA (special CB1 agonists), AM281 (CB1 antagonist), JWH133 (CB2 antagonist), SB203580 (p38 inhibitor) were from TOCRIS/R&D (Minneapolis, MN, USA). NAC and PTX were from Sigma-Aldrich (St. Louis, MO, USA). Fibronectin was from Calbiochem (Germany). YM254890 was from Adipogen Corp. (San Diego, CA, USA). SYTOX Green Nucleic Acid Stain was from Molecular Probes, Inc. (Eugene, OR, USA).
8
Endothelin-1 Signaling Pathway Analysis
9
Endothelin-1 Signaling Pathway Analysis
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Dulbecco’s modified Eagle’s medium (DMEM), Hank’s Balanced Salt Solution, and Penicillin-Streptomycin were from Gibco. Fetal Bovine Serum (FBS), 0.05% Trypsin, and Dulbecco’s Phosphate Buffered Saline (DPBS) were from Corning. Lipofectamine 2000 was from Invitrogen. Firefly D-Luciferin was from NanoLight Technology. Endothelin 1 (ET-1) was from Tocris. YM-254890 was from AdipoGen Life Sciences. Enzyme free cell dissociation solution, Thrombin, and Bovine Serum Albumin were from Millipore Sigma. FluoroBrite DMEM was from Thermo Fischer Scientific.
10
Cell Culture and Fluorescence Assay
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