The following were obtained from the respective suppliers: Dulbecco’s modified Eagle medium containing 25 mM HEPES, penicillin G potassium salt, streptomycin sulfate, dexamethasone, fatty acid-free bovine serum albumin, and recombinant human insulin (Sigma-Aldrich Corp., St. Louis, MO, USA); L-ascorbic acid phosphate magnesium salt n-hydrate, 3-isobutyl-1-methylxanthine (IBMX), and Triglyceride E-Test Kits (Wako Pure Chemical Industries Ltd., Osaka, Japan); fetal bovine serum (FBS) (MP Biomedicals, Solon, OH, USA); PGD2, 11d-11m-PGD2, MRE-269, BW245C, and 15R-15-methyl-PGD2 (15R-15m-PGD2) (Cayman Chemical (Ann Arbor, MI, USA); M-MLV reverse transcriptase (Ribonuclease H minus, point mutant) and polymerase chain reaction (PCR) Master Mix (Promega Corp., Madison, WI, USA).
Mre 269
Manufactured by Cayman Chemical
Sourced in United States
MRE-269 is a laboratory product manufactured by Cayman Chemical. It is a molecular reagent used in scientific research applications. The core function of MRE-269 is to facilitate chemical analysis and experimentation within a laboratory setting.
Automatically generated - may contain errors
Lab products found in correlation
Cay10441, by Cayman Chemical (4 mentions)
Recombinant human insulin, by Merck Group (1 mentions)
Fatty acid free bovine serum albumin, by Merck Group (1 mentions)
3 isobutyl 1 methylxanthine, by Fujifilm (1 mentions)
L ascorbic acid phosphate magnesium salt n hydrate, by Fujifilm (1 mentions)
Triglyceride e test kit, by Fujifilm (1 mentions)
Bw245c, by Cayman Chemical (1 mentions)
Pcr master mix, by Promega (1 mentions)
M mlv reverse transcriptase, by Promega (1 mentions)
Endothelin 1 peptide, by Enzo Life Sciences (1 mentions)
Pf 04418948, by Bio-Techne (1 mentions)
Treprostinil, by Bio-Techne (1 mentions)
Pdgf bb, by Merck Group (1 mentions)
Iloprost, by Merck Group (1 mentions)
L165 041, by Calibre Scientific (1 mentions)
Ns 398, by Cayman Chemical (1 mentions)
Rat anti mouse cd45, by BD (1 mentions)
Rat anti mouse f4 80, by Bio-Rad (1 mentions)
Tranylcypromine, by Enzo Life Sciences (1 mentions)
Gw1100, by Cayman Chemical (1 mentions)
8 protocols using mre 269
1
Adipocyte Differentiation Culture Protocol
2
Pharmacological Modulation of Prostanoid Receptors
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MRE-269 ([4[(5,6-diphenylpyrazinyl)(1methylethyl) amino]butoxy]acetic acid was purchased from Cayman Chemical Company (Ann Arbor, MI, USA) and PF-04418948 (selective EP2 antagonist) was purchased from Tocris Bioscience (Bristol, UK). RO-1138452 (IP selective antagonist) and butaprost (15-deoxy-16S-hydroxy-17-cyclobutyl PGE1 methyl ester), a selective EP2 agonist, was purchased from Cambridge Bioscience UK and endothelin-1 peptide from Enzo Life Science (Exeter, UK). Treprostinil was supplied by the United Therapeutics Corp (Research Triangle Park, NC, USA). Stocks of all drugs were made up in sterile DSMO (Sigma-Aldrich) to a final concentration of 10 mM. Drugs were serially diluted in growth medium, with the solvent concentration in each well remaining constant (0.1%).
3
Comprehensive Pharmacological Evaluation
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All drugs for the following investigation were procured from Tocris Bioscience (Oxford, UK). Excluding CAY‐10441 and MRE‐269, which were acquired from Cayman chemical (Michigan, USA), Rp‐8‐Br‐cAMP which was aquired from Sigma‐Aldrich (UK) and M119K which was provided by the National Cancer Institute Drug Development Programme. All drugs were dissolved in DMSO and final vehicle concentrations were always ≤0.01. For materials regarding morpholino transfection, immunodetection or RT‐qPCR, see relevant sections above.
4
Exploring Prostacyclin Analogs in Cellular Signaling
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Iloprost and PDGF-BB were purchased from Merck, MRE-269 was purchased from Cayman Chemical and treprostinil was purchased from Tocris.
5
Prostaglandin Analog-Mediated Macrophage Activation
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asc-MDM, ascTAM, ascTU or CAF were serum-deprived for 24 h in serum-free OCMI basal medium prior to stimulation with PGI2 analog MRE-269 (selexipag-active metabolite, Cayman Chemicals), Iloprost or Treprostinil (both Sigma Aldrich) for the indicated time points and concentrations. We have chosen these analogs due to different affinities to PTGIR and prostaglandin receptors, with MRE-269 as the most specific for PTGIR [26 (link)]. In individual experiments, cells were pretreated with 1 µM PTGIR antagonist CAY10449 or CAY10441 (both Cayman Chemicals) for 1 h before addition of PGI2 analog. PPARβ/δ agonist L165041 (Biozol) was applied at 1 µM concentrations where indicated. For generation of CM, asc-MDM or ex vivo TAM were stimulated with PGI2 analog under serum-free conditions for 0 or 24 h, 37 °C, 5% CO2.
6
Pharmacological Excitotoxic Model in Brain
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(RS)-(Tetrazol-5-yl)glycine (5 μM, TZG; Tocris Bioscience, Ellisville, MO, USA), a potent N-Methyl-D-aspartate (NMDA) agonist, was dissolved in phosphate-buffered saline (PBS) (pH 7.4; Thermo Fisher Scientific, Waltham, MA, USA) to induce pharmacological excitotoxic lesions in the brain. NS-398 (2 mg/kg; Cayman Chemical Company, Ann Arbor, MI, USA) and MRE-269 (1 mg/kg; Cayman Chemical Company) were dissolved in dimethyl sulfoxide before being further diluted in PBS. Tranylcypromine (10 mg/kg; Enzo Life Sciences Inc., Farmingdale, NY, USA) was dissolved in PBS (pH 7.4). Antibodies used for immunohistochemistry were rabbit anti-mouse PGI2 synthase (PGIS) (Abcam, Cambridge, MA, USA), rat anti-mouse CD45 (BD Biosciences, San Jose, CA, USA), biotinylated anti-mouse Ly6G (BD Biosciences), and rat anti-mouse F4/80 (ABD Serotec, Raleigh, NC, USA). Antibodies used for flow cytometry were fluorescein isothiocyanate-conjugated anti-Ly6G and peridinin chlorophyll protein-conjugated anti-Gr1 (BD Biosciences).
7
Comparative Analysis of Prostaglandin Receptor Modulators
8
Analyzing Prostacyclin Signaling Pathways
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Culture media and buffer solutions were purchased from Lonza Verviers (Verviers, Belgium), fetal bovine serum, insulin and trypsin from Invitrogen (CergyPontoise, France), hFGF2 from Peprotech (Neuilly sur Seine, France). cPGI2, CAY10441, GW9662, MRE269 and rosiglitazone were purchased from Cayman (BertinPharma, Montigny le bretonneux, France). All the other products, when not indicated, were from Sigma-Aldrich (Saint Quentin Fallavier, France).
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