Er50891

Manufactured by Bio-Techne

Sourced in Spain

The ER50891 is a laboratory equipment product manufactured by Bio-Techne. It is designed to perform a specific function in a research or testing environment. The core function of this product is to [description not available].

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Lab products found in correlation

7 protocols using er50891

Retinoid Receptor Agonists and Antagonists

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The following RAR and RXR agonists and antagonists were used: general RAR agonist TTNPB (4-[(E)-2-(5,6,7,8-Tetrahydro-5,5,8,8-tetramethyl-2-naphthalenyl)-1-propenyl] benzoic acid; Sigma-Aldrich) [26 (link)]; general RXR agonist methoprene acid (2E,4E-11-Methoxy-3,7,11-trimethyl-2E,4E-dodecadienoic acid; Enzo Life Sciences) [27 (link); 28 (link)]; specific RARα antagonist (ER 50891; Tocris Bioscience) [29 (link)] and specific RARß antagonist (LE 135; Tocris Bioscience) [30 (link)].
Retinoids were prepared as 10 mM (agonists) or 2.5 mM (antagonists) stock solutions in DMSO. WERI-Rb1 cells were treated for 48 h with 5 μM TTNPB or methoprene acid and with 10 μM of the RAR antagonists LE135 and ER50891, then fixed with 4% paraformaldehyde for 1h at 4°C and stained with DAPI. Control cells were treated for 48 h with an equivalent amount of DMSO, the solvent used for the agonists and antagonists.

Müller P., Doliva R., Busch M., Philippeit C., Stephan H, & Dünker N. (2015). Additive Effects of Retinoic Acid (RA) and Bone Morphogenetic Protein 4 (BMP-4) Apoptosis Signaling in Retinoblastoma Cell Lines. PLoS ONE, 10(7), e0131467.

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Adipogenic Differentiation and Lipid Quantification

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3T3-L1 pre-adipocytes were maintained and differentiated for 8 days (or 16 days where stated) as previously described [6] (link). C3H10T1/2 cells were similarly treated however penicillin/streptomycin was omitted from the media. DMSO was used as vehicle control (VEH) and to dissolve all experimental compounds. FEN (Cilag AG, Schaffhausen, Switzerland), RA (Sigma–Aldrich, UK) and ROSI (Cayman Chemical, MI, USA) were used at 1 μM, 4-OXO (Santa Cruz, TX, USA) at 0.5 μM and ER50891 (Tocris Bioscience, Bristol, UK) at 10 μM and added at day 0 (or day 8 where indicated). Cells were stained for neutral lipids with Oil Red O, images taken and then the stain was eluted and quantified at 520 nm.

Mcilroy G.D., Tammireddy S.R., Maskrey B.H., Grant L., Doherty M.K., Watson D.G., Delibegović M., Whitfield P.D, & Mody N. (2016). Fenretinide mediated retinoic acid receptor signalling and inhibition of ceramide biosynthesis regulates adipogenesis, lipid accumulation, mitochondrial function and nutrient stress signalling in adipocytes and adipose tissue. Biochemical Pharmacology, 100, 86-97.

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Diverse Pharmacological Modulators of Nuclear Receptors

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AM580 (Tocris, #0760, Germany), ER50891 (Tocris, #3823, Germany), CD2314 (Tocris, #3824, Germany), LE135 (Tocris, #2021, Germany), CD437 (Sigma, #C5865, USA), MM11253 (Tocris, #3822, Germany), ATRA (Sigma, #R2625, USA), BMS493 (Sigma, #B6688, USA), GW7647 (Tocris, #1677, Germany), GW6471 (Tocris, #4618, Germany), GW0742 (Tocris, #2229, Germany), GSK3787 (Tocris, #3961, Germany), Troglitazone (Sigma, #T2573, USA), T0070907 (Sigma, #T8703, USA), T0901317 (Sigma, #T2320, USA), SR9238 (Tocris, #5854, Germany), GC1 (Tocris, #4554, Germany), Calcifediol (Tocris, #4036, Germany), SR9011 (Sigma, #SML2067, USA), SR8278 (Tocris, #4463, Germany), CD3254 (Tocris, #3302, Germany), HX531 (Sigma, #SML2170, USA), Testosterone (Aladdin, #T102169, China), Nilutamide (Sigma, #N8534, USA), XCT790 (Sigma, #X4753, USA), Dexamethasone (Sigma, #D4902, USA), Mifepristone (Sigma, #M8046, USA), Corticosterone (Aladdin, #C104537, China), Eplerenone (Sigma, #E6657, USA), Doxorubicin (Solarbio, #D8740, China), Blasticidin (Beyotime, #ST018, China), Puromycin (Beyotime, # ST551, China).

Yuan B., Shi K., Zha J., Cai Y., Gu Y., Huang K., Yue W., Zhai Q., Ding N., Ren W., He W., Xu Y, & Wang T. (2023). Nuclear receptor modulators inhibit osteosarcoma cell proliferation and tumour growth by regulating the mTOR signaling pathway. Cell Death & Disease, 14(1), 51.

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Generating DAPT-resistant Cell Lines

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A list of the characteristics, origin and growth conditions of the cell lines used in the study is available in Supplementary Methods. To obtain DAPT-resistant cell lines, MB-157 cells were cultured in the presence of the γ-secretase inhibitor (1 μM) for 40 days. Surviving cells were diluted in medium and replated at low density to isolate single-cell derived colonies. These cell cultures were grown in medium containing DAPT (1 μM) for 83 days. Two growing clones were isolated to obtain an equivalent number of DAPT-resistant cell lines (MB-157RCL7A and MB-157RCL15A). MB-157 cells stably over-expressing RARβ-targeting shRNAs were obtained by lentiviral infection of constructs based on the pGreenPuro-shRNA system (SBI, System Bioscences). The sequence of the RARβ-targeting shRNAs is available in Supplementary Methods. Lentiviral infected cells were subjected to puromycin (0.5 µg/mL) selection for the isolation of the shRARβ expressing cells.
The sensitivity of cell lines to the anti-proliferative action of ATRA was evaluated with the ATRA-score index [17 (link),18 (link)]. The following chemicals were used: ATRA (Sigma-Aldrich), AM580 (Tocris), BMS961 (Tocris), UVI2003 (a kind gift of Dr. Angel De Lera, Universidade de Vigo, Spain), ER50891 (Tocris), DAPT (Sigma), PF-03084014 (Pfeizer) and VP16 (SIGMA).

Paroni G., Zanetti A., Barzago M.M., Kurosaki M., Guarrera L., Fratelli M., Troiani M., Ubezio P., Bolis M., Vallerga A., Biancardi F., Terao M, & Garattini E. (2020). Retinoic Acid Sensitivity of Triple-Negative Breast Cancer Cells Characterized by Constitutive Activation of the notch1 Pathway: The Role of Rarβ. Cancers, 12(10), 3027.

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Retinoid Receptor Modulation in Breast Cancer

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The following compounds were used: ATRA (Sigma-Aldrich, https://www.sigmaaldrich.com), AM580 (Tocris, http://www.tocris.com), BMS961 (Tocris), ER50891 (Tocris), CD2665 (Tocris), and UVI2003 (a kind gift of Dr. Angel De Lera, Universidade de Vigo, Spain). Sulforhodamine was from Sigma-Aldrich Co. A list of the cell lines, their characteristics, and origin is available in Supplementary Table S1. The plasmid constructs used for RARα3 over-expression in MDA-MB453 cells and knockdown in SKBR3 cells are described below.

Centritto F., Paroni G., Bolis M., Garattini S.K., Kurosaki M., Barzago M.M., Zanetti A., Fisher J.N., Scott M.F., Pattini L., Lupi M., Ubezio P., Piccotti F., Zambelli A., Rizzo P., Gianni' M., Fratelli M., Terao M, & Garattini E. (2015). Cellular and molecular determinants of all-trans retinoic acid sensitivity in breast cancer: Luminal phenotype and RARα expression. EMBO Molecular Medicine, 7(7), 950-972.

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Signaling Pathways in Cell Differentiation

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Recombinant human IFN-γ was purchased from the R&D systems. Recombinant human LT-α2β1 (#679-TX-010), murine LT-α2β1 (#1008-LY-010), human IL-1β, human IL-22, human Wnt-3a, and anti-LT-β receptor antibody (#AF629) were also purchased from the R&D systems. Recombinant human RANK ligand was purchased from PeproTech (#310-01). Retinoic acid (#695) and RAR inhibitors ER50891, LE135, MM11253, BMS-493 were purchased from Tocris. Retinol and tamoxifen were acquired from Sigma-Aldrich and RARα agonist AM580 was purchased from Wako (Alpha Laboratories). Red fluorescence labeled carboxylate-modified polystyrene latex beads with 0.5 μm diameter were purchased from Sigma-Aldrich. Antibody used for RelB chromatin immunoprecipitation (sc-226) was purchased from Santa Cruz Biotechnology.

Ding S., Song Y., Brulois K.F., Pan J., Co J.Y., Ren L., Feng N., Yasukawa L.L., Sánchez-Tacuba L., Wosen J.E., Mellins E.D., Monack D.M., Amieva M.R., Kuo C.J., Butcher E.C, & Greenberg H.B. (2020). Retinoic Acid and Lymphotoxin Signaling Promote Differentiation of Human Intestinal M Cells. Gastroenterology, 159(1), 214-226.e1.

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Retinoic Acid Reconstitution Protocol

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RA was purchased from Sigma (cat#R2625) and reconstituted in DMSO at a stock concentration of 100mM (in vitro experiments) or 40mg/mL (in vivo experiments). BMS753 (Tocris cat#3505) and ER50891 Tocris (cat#3823) were reconstituted in DMSO to stock concentrations of 100mM. Aliquots of RA and agonists/antagonists were serially diluted in PBS immediately prior to in vitro treatment of cells. Stocks were replaced every 3–6 months.

Schwartz D.M., Farley T.K., Richoz N., Yao C., Shih H.Y., Petermann F., Zhang Y., Sun H.W., Hayes E., Mikami Y., Jiang K., Davis F.P., Kanno Y., Milner J.D., Siegel R., Laurence A., Meylan F, & O’Shea J.J. (2019). Retinoic acid receptor alpha represses a Th9 transcriptional and epigenomic program to reduce allergic pathology. Immunity, 50(1), 106-120.e10.

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