T0901317

Manufactured by Cayman Chemical

Sourced in United States

T0901317 is a synthetic small molecule that functions as an agonist for the liver X receptor (LXR) family of nuclear receptors. This compound is commonly used in scientific research to study the biological effects mediated by LXR activation.

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Lab products found in correlation

Gw3965, by Merck Group (3 mentions) Gw3965, by Cayman Chemical (3 mentions) Lipopolysaccharides (lps), by Merck Group (3 mentions) Oil red o, by Merck Group (3 mentions) Streptomycin, by Thermo Fisher Scientific (2 mentions) Dimethyl sulfoxide (dmso), by Merck Group (2 mentions) Pioglitazone, by Merck Group (2 mentions) β actin, by Cell Signaling Technology (2 mentions) Rosiglitazone, by Cayman Chemical (2 mentions) Bexarotene, by LC Laboratories (2 mentions) Lipofectamine rnaimax, by Thermo Fisher Scientific (2 mentions) Penicillin, by Thermo Fisher Scientific (1 mentions) Ethanol, by Avantor (1 mentions) Brassicasterol, by Merck Group (1 mentions) Fucosterol, by Merck Group (1 mentions) Stigmasterol, by Cayman Chemical (1 mentions) Fetal calf serum (fcs), by Thermo Fisher Scientific (1 mentions) β sitosterol, by Merck Group (1 mentions) β sitosterol, by Cayman Chemical (1 mentions) Dulbecco modified eagle medium (dmem), by Merck Group (1 mentions)

52 protocols using t0901317

Modulation of NR1H3 promoter activity

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Hep3B and COS-7 cells were transfected with NR1H3 promoter pGL3-basic constructs using FuGENE6 (Promega) according to the manufacturer’s instructions. After incubation for six hours, the medium was replenished with 500 μL of fresh medium with 20% FBS, and the cells were incubated a further 18 hours at 37°C in a 5% CO2 incubator. Twenty four hours after transfection, cells were treated with either 200 ng/mL LPS (Sigma-Aldrich, St. Louis, MO, USA), 3 μmol/L 3-[3-[N-(2-chloro-3-trifluoro methylbenzyl)-(2,2-diphenylethyl)amino]propyloxy]phenylacetic acid hydrochloride (hydrochloride GW3965; GW3965, Sigma-Aldrich) or 5 μmol/L N-(2,2,2-trifluoroethyl)-N-[4-[2,2,2-trifluoro-1-hydroxy-1-(trifluoromethyl)ethyl]phenyl]-benzenesulfonamide (sulfonamide T0901317; T0901317, Cayman, Ann Arbor, MI, USA) for 24 hours. These concentrations and time were chosen based on previous work
[16 (link)].

Jeon J.Y., Nam J.Y., Kim H.A., Park Y.B., Bae S.C, & Suh C.H. (2014). Liver X receptors alpha gene (NR1H3) promoter polymorphisms are associated with systemic lupus erythematosus in Koreans. Arthritis Research & Therapy, 16(3), R112.

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NASH Mice Model with CCl4 and T0901317

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NASH mice were fed an HF diet (60 kcal% fat; D12492, Research Diets, Inc., New Brunswick, NJ, United States) for 4 wk, intraperitoneally injected with CCl₄ (Wako Pure Chemical Industries, Ltd., Osaka, Japan) twice a week for the final 2 wk, and intraperitoneally injected with T0901317 (Cayman Chemical Co., Ann Arbor, MI, United States) solubilized in DMSO for the final 5 d. The CCl₄ dose was 0.1 mL/kg, and the T0901317 dose was 2.5 mg/kg[28 (link)].

Ozawa Y., Tamura T., Owada Y., Shimizu Y., Kemmochi A., Hisakura K., Matsuzaka T., Shimano H., Isoda H, & Ohkohchi N. (2018). Evaluation of safety for hepatectomy in a novel mouse model with nonalcoholic-steatohepatitis. World Journal of Gastroenterology, 24(15), 1622-1631.

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Phytosterol Effects on Cell Lines

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Immortalized human embryonic kidney cells (HEK293.T), human microglia (CHME3; a kind gift from prof. dr. M. Tardieu, Universite Paris-Sud, France^{66 (link)}), human oligodendrocytes (MO3.13), mouse neuroblastoma expressing APPswe (N2a/APPswe; a kind gift from prof. dr. T.W. Kim, Colombia University, USA^{67 (link)}), and monkey kidney cells (COS7) were used for in vitro experiments. All cell lines were cultured in DMEM (Sigma-Aldrich) containing 10% heat-inactivated FCS (Invitrogen, Merelbeke, Belgium) and 100 U penicillin/100 µg streptomycin/ml (Invitrogen), at 37 °C/5% CO₂. For phytosterol treatment, cells were incubated for 18 hours in culture medium without FCS containing the Eastern plants extracts, brassicasterol (Sigma-Aldrich), β-sitosterol, (Sigma-Aldrich), fucosterol (Sigma-Aldrich), stigmasterol (analytic confirmed purity of 99,9%), phytosterol mix (containing 60% β-sitosterol, 25% campesterol, and 15% stigmasterol; kindly provided by Ingmar Wester Raisio, Finland), T0901317 (Cayman Chemicals, Huissen, the Netherlands), ethanol (VWR), or DMSO (Sigma-Aldrich).

Bogie J., Hoeks C., Schepers M., Tiane A., Cuypers A., Leijten F., Chintapakorn Y., Suttiyut T., Pornpakakul S., Struik D., Kerksiek A., Liu H.B., Hellings N., Martinez-Martinez P., Jonker J.W., Dewachter I., Sijbrands E., Walter J., Hendriks J., Groen A., Staels B., Lütjohann D., Vanmierlo T, & Mulder M. (2019). Dietary Sargassum fusiforme improves memory and reduces amyloid plaque load in an Alzheimer’s disease mouse model. Scientific Reports, 9, 4908.

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Dietary Modulation of Lipid Metabolism

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C57BL/6 J male and female mice were purchased from Jackson Laboratory (Bar Harbor, ME) at 6 weeks of age and housed in individually ventilated cages (14:10, light: dark) with free access to feed and water. Test diets were formulated from control diet (Research Diets Inc., AIN-76A) and supplemented with either 0.3% (w/w) stigmasterol (Sigma-Aldrich, Product Number S2424) or 0.015% (w/w) T0901317 (Cayman Chemical, Product Code 71810). At 8 weeks of age, mice were singly housed with access to water and control diet for 3 days. On day four, mice were randomized to receive either control diet, stigmasterol-, or T0901317-supplemented diet (ad libidum) for an additional 4 days. On Day 8, mice were euthanized for plasma and tissue dissection (n=8 per sex). Mice were euthanized by carbon dioxide and exsanguination via cardiac puncture at termination. Liver and small intestine sections were dissected, snap frozen in liquid nitrogen, and stored in −80 °C. Small intestine was divided into three equal sections representing the duodenum, jejunum, and ileum. Blood was collected in EDTA-coated tubes, centrifuged at 2000×g for 15 min at 4 °C, and plasma stored at −80 °C.

Lifsey H.C., Kaur R., Thompson B.H., Bennett L., Temel R.E, & Graf G.A. (2019). Stigmasterol stimulates transintestinal cholesterol excretion independent of liver X receptor activation in the small intestine. The Journal of nutritional biochemistry, 76, 108263.

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Immunoblotting Assay for AMPK Pathway

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Anti-AMPKα, anti-phosphor-AMPKα (Thr172), anti-ACC, anti-phosphor-ACC (Ser 79) and anti-SCD1 antibodies were purchased from Cell Signaling Technology, Inc. Anti-FAS, anti-MTP, anti-PPARα and anti-LXRα antibodies were obtained from Abcam (Cambridge, MA, USA). T0901317 and GW9508 were purchased from Cayman Chemical (Ann Arbor, MI). STO-609 was purchased from Sigma-Aldrich (St. Louis, MO, USA).

Li M., Meng X., Xu J., Huang X., Li H., Li G., Wang S., Man Y., Tang W, & Li J. (2016). GPR40 agonist ameliorates liver X receptor-induced lipid accumulation in liver by activating AMPK pathway. Scientific Reports, 6, 25237.

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LXRα Agonist T0901317 Assay

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The LXRα agonist T0901317 was obtained from Cayman Chemical (Ann Arbor, MI, USA). The primary Ab directed at LXRα was obtained from Abcam (Cambridge, MA, USA). The secondary Abs and β-actin were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). All other chemicals were obtained from Sigma-Aldrich (St. Louis, MO, USA).

Botez G., Piraino G., Hake P.W., Ledford J.R., O’Connor M., Cook J.A, & Zingarelli B. (2015). Age-dependent therapeutic effects of liver X receptor-α activation in murine polymicrobial sepsis. Innate Immunity, 21(6), 609-618.

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T0901317, Pioglitazone, and GW3965 Preparation

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T0901317 was purchased from Cayman Chemical Company (Ann Arbor, Michigan, USA). Pioglitazone and GW3965 were purchased from Sigma-Aldrich (Saint Louis, Missouri, USA). In animal experiments, T0901317 was solubilized in a vehicle containing 3% dimethyl sulfide (DMSO) in PBS and administered by intraperitoneal injection at a dose of 30 mg/kg body weight. In cellular experiments, T0901317 and GW3965 were prepared in a solution of 1∶1 DMSO: PBS at a concentration of 1 mM, and Pioglitazone was solubilized in DMSO at a concentration of 20 mM for further dilution with cell medium.

Zheng F., Zhang S., Lu W., Wu F., Yin X., Yu D., Pan Q, & Li H. (2014). Regulation of Insulin Resistance and Adiponectin Signaling in Adipose Tissue by Liver X Receptor Activation Highlights a Cross-Talk with PPARγ. PLoS ONE, 9(6), e101269.

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Synthesis and Dilution of LXR Agonist and Antagonist

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A synthetic LXR agonist, T0901317 (TO), was purchased from Cayman Chemical (Ann Arbor, MN, USA), and a synthetic LXR antagonist, GSK2033 (GSK), was purchased from Axon Medchem BV (Groningen, The Netherlands). TO and GSK were dissolved in and diluted with dimethyl sulfoxide at various concentrations.

Sakane Y., Kanamoto N., Yamauchi I., Tagami T., Morita Y., Miura M., Sone M., Yasoda A., Kimura T., Nakao K, & Inagaki N. (2017). Regulation of type 1 iodothyronine deiodinase by LXRα. PLoS ONE, 12(6), e0179213.

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Protein Expression and Antibody Validation

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T0901317 was purchased from Cayman Chemical (Ann Arbor, Michigan, USA). Gefitinib (Iressa) was purchased from AstraZeneca UK Limited (Macclesfield, UK). All drugs were suspended in DMSO and stored at −20°C. Antibodies against MMPs, E-cadherin, and β-actin were obtained from Cell Signaling Technology (Danvers, Massachusetts, USA).

Lou R., Cao H., Dong S., Shi C., Xu X., Ma R., Wu J, & Feng J. (2019). Liver X receptor agonist T0901317 inhibits the migration and invasion of non-small-cell lung cancer cells in vivo and in vitro. Anti-Cancer Drugs, 30(5), 495-500.

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Synthesis and Characterization of Oxysterol Compounds

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27HC (purity >95%) and dendrogenin A (purity >98%) were synthesized by Sai Life (Hyderabad, India). 25HC, 4βHC and Desmosterol were synthesized by Steraloids (Newport, RI) (purity >98%). 22(R)HC was synthesized by Avanti (Alabaster, AL) (purity >99%). 24(S)HC and 24,25-epoxycholesterol (24,25EC) were synthesized by Enzo (Farmingdale, NY) (purity ≥98%). GW3965, T0901317, GSK2033 and 7-ketocholesterol (7KC) were synthesized by Cayman (Ann Arbor, MI) (purity ≥98%). 20(S)HC was synthesized by Tocris (Bristol, UK) (purity >95%). 17β-Estradiol (E2) was synthesized by MP Biomedicals (Irvine, California) (purity ≥98%). 27HC, dendrogenin A, 24(S)HC, 20(S)HC, GW3965, T0901317 and GSK2033 were dissolved in dimethyl sulfoxide (DMSO) and stored in −20°C. 7KC, 4βHC, 22(R)HC, 24,25EC and 25HC were dissolved in ethanol and stored in −20°C. Desmosterol was dissolved in M-pyrol and stored in −20°C. Antibodies for flow cytometry were purchased from BD Biosciences and used at a working dilution of 1:100 in FACS buffer (2% fetal bovine serum (FBS) in phosphate buffered saline (PBS)). Catalogue numbers were as follows: CD3: 555275, CD4: 553729, CD8: 557682. Annexin V-FITC/propidium iodide apoptosis assay was purchased from Thermo Fisher (catalogue number: V13242), and FAM-DEVD-FMK/propidium iodide apoptosis assay was purchased from Immunochemistry (catalogue number: 94).

Ma L., Wang L., Nelson A.T., Han C., He S., Henn M.A., Menon K., Chen J.J., Baek A.E., Vardanyan A., Shahoei S.H., Park S., Shapiro D.J., Nanjappa S.G, & Nelson E.R. (2020). 27-Hydroxycholesterol acts on myeloid immune cells to induce T cell dysfunction, promoting breast cancer progression.. Cancer letters, 493, 266-283.

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