Gw3965

Manufactured by Selleck Chemicals

Sourced in United States

GW3965 is a synthetic agonist of the liver X receptor (LXR) transcription factor. It functions by activating LXR, which is involved in the regulation of genes related to cholesterol and lipid metabolism.

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GW3965 HCL (2 citations)

8 protocols using gw3965

Gingival Response to LXR and PPAR Agonists

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18-week-old CD18^+/− and CD18^−/− mice were locally microinjected with a combination of GW3965 and GW0742 (both at 8.1 nmol; Selleckchem) or dimethyl sulfoxide (DMSO) vehicle control. GW3965 is a selective ligand for LXRs although it cannot distinguish between LXRα and LXRβ (EC_50s of 190 and 30 nM, respectively) [44 (link)]. GW0742 is a selective agonist of PPARβ/δ with EC₅₀ value of 2 nM; although it can exhibit agonistic activity also for PPARγ, the corresponding EC₅₀ value is much higher (2.4 μM) [45 (link)]. These agonists were administered using a split-mouth experimental design; i.e., one side was locally injected in the palatal gingiva of the 2^nd molar with the drug combination and the contralateral side with vehicle control. After 72h, the mice were euthanized. Dissected gingiva were processed for quantitative real-time PCR (section 2.6) and defleshed maxilla were used to measure bone heights (section 2.2).

Kajikawa T., Wang B., Li X., Wang H., Chavakis T., Moutsopoulos N.M, & Hajishengallis G. (2020). Activation of metabolic nuclear receptors restores periodontal tissue homeostasis in mice with leukocyte adhesion deficiency-1. Journal of leukocyte biology, 108(5), 1501-1514.

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Modulating Neurogenesis with LXR Agonist GW3965

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In the CUS/GW3965 group, the animals were intraperitoneally injected with the LXR agonist GW3965 (Selleckchem, S2630, 10 mg/kg/day) for 28 consecutive days (Collins et al., 2002 (link); Morales et al., 2008 (link)). The Control/standard group and the CUS/standard group were intraperitoneally injected with the same volume of normal saline (Figure 1A). During the periods of administration, the mice in the CUS/standard group and the CUS/GW3965 group were subjected to the stressors. In the present experiment, BrdU was diluted to a concentration of 10 mg/ml in normal saline. All the animals were intraperitoneally injected with BrdU at a dosage of 50 mg/kg/day for 11 consecutive days starting in the sixth week. The concentration and dosage adopted to evaluate the effect of GW3965 on cell proliferation were referred to the previous studies (Wojtowicz and Kee, 2006 (link); HG and CM, 2007 (link); Qiu et al., 2007 (link); Mateus-Pinheiro et al., 2013 (link); Tang et al., 2021 (link); Liang et al., 2022 (link)).

Zhu P., Tang J., Liang X., Luo Y., Wang J., Li Y., Xiao K., Li J., Deng Y., Jiang L., Xiao Q., Qi Y., Xie Y., Yang H., Zhu L., Tang Y, & Huang C. (2022). Activation of liver X receptors protects oligodendrocytes in CA3 of stress-induced mice. Frontiers in Pharmacology, 13, 936045.

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NDV Protein Detection Protocol

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GW3965 was purchased from Selleck (China). Cholesterol-water soluble was purchased from Sigma. Mouse monoclonal β-actin antibody (Sigma), goat anti-mouse IgG (Beyotime), and goat anti-rabbit IgG (Beyotime, China) were used in the studies. Filipin III was purchased from Cayman Chemical (U.S.A.). A mouse monoclonal antibody against NDV nucleocapsid protein (NP) was prepared in our laboratory.

Sheng X.X., Sun Y.J., Zhan Y., Qu Y.R., Wang H.X., Luo M., Liao Y., Qiu X.S., Ding C., Fan H.J, & Mao X. (2016). The LXR ligand GW3965 inhibits Newcastle disease virus infection by affecting cholesterol homeostasis. Archives of Virology, 161(9), 2491-2501.

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Modulation of Lipid Metabolism Pathways

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Reagents Sorafenib (multikinase inhibitor), T0901317 (LXR pan-agonist), GW3965 (LXR pan-agonist), PF-04217903 (ATP-competitive Met inhibitor), Gefitinib (EGFR-tyrosine kinase inhibitor), MK-2206 (Akt1/2/3 inhibitor), SCH772984 (ERK1/2 inhibitor), and SB202190 (p38 MAPK Inhibitor) were purchased from Selleck Chemicals (Houston, TX, US). Antibodies against LXRα and LXRβ were purchased from Abcam cooperation (Cambridge, UK). All other antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA). Puromycin and TRIzol reagent were purchased from Thermo Fisher Scientific (Grand Island, NY, USA). Cell Counting Kit-8 (CCK-8) was purchased from Dojindo Laboratories (Mashikimachi, kamimashiki gun Kumamoto, JAPAN). PI/RNase Staining Buffer and FITC Annexin V Apoptosis Detection Kit were purchased from BD Biosciences (San Diego, CA, USA). Cholesterol Assay Kit was purchased from Invitrogen (San Diego, CA, USA). BCA Protein Assay Reagent was purchased from Beyotime Biotechnology (Shanghai, China).

Shao W., Zhu W., Lin J., Luo M., Lin Z., Lu L., Jia H., Qin L., Lu M, & Chen J. (2019). Liver X Receptor Agonism Sensitizes a Subset of Hepatocellular Carcinoma to Sorafenib by Dual-Inhibiting MET and EGFR. Neoplasia (New York, N.Y.), 22(1), 1-9.

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Rat Cardiomyocyte Cell Line H9C2 Culture

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The rat cardiomyocyte cell line H9C2 was obtained from ScienCell Research Laboratories, Inc. (San Diego, CA, USA) and cultured in Dulbecco's modified Eagle's medium (DMEM; Gibco; Thermo Fisher Scientific, Inc., Waltham, MA, USA) with 10% heat-inactivated fetal bovine serum (FBS; Hyclone; GE Healthcare Life Sciences, Logan, UT, USA) and 100 U/ml penicillin and streptomycin (Gibco; Thermo Fisher Scientific, Inc.). Cells were maintained at 37°C in a humidified atmosphere containing 5% CO₂. GW3965, an LXR agonist, was purchased from Selleck Chemicals (Houston, TX, USA) and glucose was purchased from Sigma-Aldrich (Merck KGaA Darmstadt, Germany).

Cheng Y., Zhao W., Zhang X., Sun L., Yang H., Wang Y., Cao Y., Chu Y, & Liu G. (2018). Downregulation of microRNA-1 attenuates glucose-induced apoptosis by regulating the liver X receptor α in cardiomyocytes. Experimental and Therapeutic Medicine, 16(3), 1814-1824.

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LXR Modulation and Metabolic Regulation

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ABT263, WEHI‐539, A1210477, GW3965, LXR623, ABT199, and adenosine triphosphate (ATP) were obtained from Selleckchem. Low‐density lipoprotein (LDL) from human plasma was acquired from Thermo Fisher. l‐Aspartic acid sodium salt monohydrate was purchased from Sigma. Reagents were dissolved in dimethylsulfoxide (DMSO) (10 mM stock solution) and stored at −20°C. Final concentrations of DMSO were below 0.1% (v/v). The following antibodies were used: ABCA1 (Abcam ab18180; 1:25), Mcl‐1 (Cell Signaling Technology (CST) 5453SS; 1:500), BAK (CST 12105S; 1:500), Bcl‐2 (Abcam ab59348; 1:500), BIM (CST 2933S; 1:500), Bcl‐xL (CST 2764S; 1:500), Usp9X (CST 15751S; 1:1,000), Noxa (Millipore OP180; 1:500), β‐actin (Sigma‐Aldrich A1978, clone AC15; 1:2,000), ATF3 (CST 33593S; 1:500), ATF4 (CST 11815S; 1:500), Vinculin (Abcam ab129002; 1:200), AMPK (CST 5831S; 1:25), pAMPK (CST 2531S; 1:25), PAPRP (CST 9532S; 1:500), caspase‐9 (CST 9502S; 1:500), GRP78 (CST 3177S; 1:25), HA (CST 3724S; 1:500), LXRβ (Abcam ab56237; 1:25), SDHB (Abcam ab14714, 1:25), OXPHOS (Abcam ab110411; 1:500), and secondary goat anti‐mouse IgG‐HRP‐linked (SC2005) and secondary goat anti‐rabbit IgG‐HRP‐linked antibodies (SC2004) were purchased from Santa Cruz Biotechnology, Inc.

Nguyen T.T., Ishida C.T., Shang E., Shu C., Torrini C., Zhang Y., Bianchetti E., Sanchez‐Quintero M.J., Kleiner G., Quinzii C.M., Westhoff M., Karpel‐Massler G., Canoll P, & Siegelin M.D. (2019). Activation of LXRβ inhibits tumor respiration and is synthetically lethal with Bcl‐xL inhibition. EMBO Molecular Medicine, 11(10), e10769.

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Induction of Macrophage Differentiation

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Authenticated human monocytic THP-1 cells were purchased from the Cell Bank, Type Culture Collection, Chinese Academy of Sciences. THP-1 cells were maintained in RPMI-1640 medium (Gibco) supplemented with 10% FBS, 0.05 mM 2-mercaptoethanol, and 1% penicillin-streptomycin at 37°C in a humidified 5% CO₂ atmosphere. THP-1 cells (2×10⁵ per well) were seeded in 12-well plates and differentiated using 50 ng/ml phorbol 12-myristate-13-acetate (PMA; CS0001, Multi Sciences) for 48 hours to obtain macrophage-like cells. Then, the cells were treated with the LXR agonists GW3965 and T0901317 (Selleck Chemicals) at 1 μM for 24 hours. For gene silencing, differentiated THP-1 cells were transfected with small interfering RNA (siRNAs) targeting NR1H3 (LXR-α), HIF1A (HIF-1α), or scramble control (GenePharma) with GP-transfect-Mate (GenePharma). Oligonucleotide sequences for gene knockdown are listed in Supplementary Table 2. All experiments were performed with mycoplasma-free cells.

Zhou L., Wang M., Guo H., Hou J., Zhang Y., Li M., Wu X., Chen X, & Wang L. (2022). Integrated Analysis Highlights the Immunosuppressive Role of TREM2+ Macrophages in Hepatocellular Carcinoma. Frontiers in Immunology, 13, 848367.

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Dietary Regulation of Lipid Metabolism

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All animal protocols and studies were performed according to guidelines from the Center for Experimental Animals at Fukuoka University. Eight-week-old male mice (n = 4) on an ob/ob background or C57BL/6J mice with a wild-type leptin gene (OB/OB) were fed an ad libitum diet (MF, Oriental Yeast, Fukuoka, Japan) with or without 0.025% (w/w) T0901317 (Sigma Aldrich, St. Louis, MO, USA) for 2 weeks, as previously described (Matsusue et al., 2014 (link)). GW3965 (Selleck, Japan) was administered with 20 mg/day/kg for 3 days by oral gavage (Laffitte et al., 2003 (link)). As a positive control for oral gavage administration, T0901317 was administered at 20 mg/day/kg for 3 days (Jakel et al., 2004 (link)). Vehicle alone was administered as a negative control (0.5% methyl-cellulose).

Aibara D., Matsusue K., Takiguchi S., Gonzalez F.J, & Yamano S. (2018). Fat-specific protein 27 is a novel target gene of liver X receptor α. Molecular and cellular endocrinology, 474, 48-56.

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