Gw6471

Manufactured by Cayman Chemical

Sourced in United States

GW6471 is a chemical compound used in research applications. It functions as a selective peroxisome proliferator-activated receptor alpha (PPARα) antagonist. The core function of GW6471 is to inhibit the activity of the PPARα receptor.

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Lab products found in correlation

Gw9662, by Cayman Chemical (5 mentions) Gw7647, by Cayman Chemical (3 mentions) Pf3845, by Cayman Chemical (2 mentions) Cremophor, by Merck Group (2 mentions) Pioglitazone, by Cayman Chemical (2 mentions) Insulin, by Merck Group (2 mentions) Gw9662, by Merck Group (2 mentions) β hydroxybutyrate, by Cayman Chemical (1 mentions) Anti nlrp3 antibody, by Adipogen (1 mentions) Ly294002, by Cell Signaling Technology (1 mentions) Sb431542, by Cayman Chemical (1 mentions) Mcc950, by Adipogen (1 mentions) Tubulin 2144, by Cell Signaling Technology (1 mentions) Enzymatic kit, by Fujifilm (1 mentions) Ng nitro l arginine methyl ester l name, by Merck Group (1 mentions) Pemafibrate, by Kowa (1 mentions) Palmitate pal, by Merck Group (1 mentions) Perhexiline maleate, by Cayman Chemical (1 mentions) Catalase, by Merck Group (1 mentions) Bezafibrate, by Cayman Chemical (1 mentions)

22 protocols using gw6471

Generation of Adipolin Antibody and Assays

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A polyclonal antibody against mouse adipolin was generated, as previously described.⁴³ (link) Anti-NLRP3 antibody was purchased from Adipogen and Novus. Anti-caspase 1 antibodies were purchased from Adipogen and Proteintech. Anti-IL1β antibodies were purchased from R&D Systems. MCC950 was purchased from Adipogen. GW6471 and SB431542 were purchased from Cayman Chemicals. LY294002 was purchased from Cell Signaling Technology. Recombinant angiotensin II protein was purchased from Sigma-Aldrich. β-hydroxybutyrate and a ketone Body assay kit were purchased from Cayman. Adenoviral vectors expressing the genes encoding β-galactosidase (Ad-β-gal) and FLAG-tagged adipolin (Ad-APL) were constructed under the control of the CMV promoter.¹³ (link)^,¹⁴ (link)

Fang L., Ohashi K., Hayakawa S., Ogawa H., Otaka N., Kawanishi H., Takikawa T., Ozaki Y., Takahara K., Tatsumi M., Takefuji M., Shimizu Y., Bando Y.K., Fujishima Y., Maeda N., Shimomura I., Murohara T, & Ouchi N. (2023). Adipolin protects against renal injury via PPARα-dependent reduction of inflammasome activation. iScience, 26(5), 106591.

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Pemafibrate Enhances FGF21 Signaling

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Pemafibrate was kindly provided by Kowa Co. Ltd (Nagoya, Japan). Mouse CD31 antibody was purchased from BD Pharmingen (San Jose, CA)(550274). Antibodies of phosphorylated eNOS (Ser-1177)(9571), eNOS (32027) and Tubulin (2144) were purchased from Cell Signaling Technology (Beverly, MA). NG-nitro-l-arginine methyl ester (L-NAME) was purchased from Sigma (St. Louis, MO) (N5751). GW6471 was purchased from Cayman Chemical (11697). Recombinant human FGF21 protein was purchased from R&D system (2539-FG-025). Plasma FGF21 levels were measured by ELISA kit (R & D system)(MF2100) [15 (link)]. Plasma adiponectin levels were determined by ELISA kit (Otsuka Pharmaceutical Co. Ltd.)(410713). Adenoviral vectors expressing mouse full-length FGF21 (Ad-FGF21) were constructed under the control of the CMV promoter [16 (link), 17 (link)]. Adenoviral vectors expressing β-galactosidase (Ad-βgal) were used as controls [18 (link)]. Lipid profiles and plasma glucose were analyzed by enzymatic kits (Wako Pure Chemical Industries, Ltd) (total cholesterol, 439–17501) (triglyceride, 290–63701) (glucose, 439–90901).

Kawanishi H., Ohashi K., Ogawa H., Otaka N., Takikawa T., Fang L., Ozaki Y., Takefuji M., Murohara T, & Ouchi N. (2020). A novel selective PPARα modulator, pemafibrate promotes ischemia-induced revascularization through the eNOS-dependent mechanisms. PLoS ONE, 15(6), e0235362.

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Fibroblast Growth Factor Protects Renal Tubule Cells from Palmitate-Induced Injury

Cited 1 time

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Mouse proximal tubule (BUMPT) cell line was obtained from Boston University School of Medicine, Boston, MA. BUMPT cells were cultured in DMEM containing 1 mmol/L d-glucose, 10% fetal bovine serum (FBS), 100 units/mL penicillin and 100 μg/mL streptomycin, and incubated in a humidified atmosphere of 5% CO₂ at 37 °C, and passaged every 2 days by trypsinization. BUMPT cells were seeded in 6-well culture plates at an initial density of 20,000 cells/well and grown for 24 h before treatment.
Palmitate (Pal, Cat No: P9767, Sigma-Aldrich) was dissolved in 50% ethanol and mixed with bovine serum albumin (BSA) to make Pal-BSA stock solution. BUMPT cells were exposed to the Pal-BSA solution to mimic the renal tubule cell damage in diabetic mice as described in our previous studies [16 (link),21 (link)]. The peroxisome proliferator-activated receptor (PPARα) inhibitor (GW6471, Cat No: 11697, Cayman Chemical) was dissolved in dimethyl sulfoxide (DMSO). After serum starvation for 24 h, BUMPT cells were exposed to GW6471 (10 μmol/L) or DMSO in FBS free medium for 8 h. Then BUMPT cells were pretreated with FGF1^ΔHBS (100 nmol/L) for 1 h, followed by exposure to 90 μmol/L Pal-BSA or BSA in the presence and absence of FGF1^ΔHBS for additional 16 h. Then cells were collected to detect PPARα expression and apoptotic indicators.

Lin Q., Chen O., Wise JP J.r., Shi H., Wintergerst K.A., Cai L, & Tan Y. (2022). FGF1ΔHBS delays the progression of diabetic nephropathy in late-stage type 2 diabetes mouse model by alleviating renal inflammation, fibrosis, and apoptosis. Biochimica et biophysica acta. Molecular basis of disease, 1868(8), 166414.

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Modulation of Peroxisome PPAR-α Pathway

Cited 2 times

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The peroxisome PPAR-α agonist bezafibrate (Cayman Chemical, MI, USA), the PPAR-α antagonist GW6471 (Cayman Chemical, MI, USA), and/or the CPT1 inhibitor perhexiline maleate (Cayman Chemical, MI, USA) were cultured with Jurkat cell in vitro at the indicated concentrations^{41 (link), 42 (link)}. Catalase (Sigma-Aldrich) was used to inhibit ROS production at 1000 U/mL^{8 (link)}. perhexiline maleate was injected intraperitoneal (i.p.) to inducible liver-specific MYC oncogene transgenic mice (MYC-ON) at 8 mg/kg in 100 µL 50/50 solution of PBS/DMSO three times per week for a total of 5 weeks^{27 (link)}.

Brown Z.J., Fu Q., Ma C., Kruhlak M., Zhang H., Luo J., Heinrich B., Yu S.J., Zhang Q., Wilson A., Shi Z.D., Swenson R, & Greten T.F. (2018). Carnitine palmitoyltransferase gene upregulation by linoleic acid induces CD4+ T cell apoptosis promoting HCC development. Cell Death & Disease, 9(6), 620.

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Serine Hydrolase Activity-Based Profiling

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The serine hydrolase activity-based protein profiling probe (ABPP probe), FP-TAMRA, was purchased from Thermo Fisher Scientific (Waltham, MA, USA). Small interfering RNAs (FlexiTube SI02736706 for fatty acid amide hydrolase (FAAH) gene knockdown and Allstars Negative Control) were purchased from Qiagen (Tokyo, Japan), and transfection reagent Lipofectamine RNAiMAX was purchased from Thermo Fisher Scientific. All inhibitors, antagonists, and reagents, including PF3845, URB597, SR141716, SR144528, GW9662, GW6471, O1918, anandamide (AEA), and arachidonic acid (AA), were purchased from Cayman Chemical (Ann Arbor, MI, USA).
Radioactive anandamide [ethanolamide-1,2-¹⁴C] was purchased from American Radiolabeled Chemicals, Inc (Saint Louis, MO, USA). Other chemicals including lipopolysaccharide were purchased from Sigma-Aldrich (St. Louis, MO, USA).

Tanaka M., Yagyu K., Sackett S, & Zhang Y. (2019). Anti-Inflammatory Effects by Pharmacological Inhibition or Knockdown of Fatty Acid Amide Hydrolase in BV2 Microglial Cells. Cells, 8(5), 491.

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Intestinal Epithelial Cell Culturing

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The human intestinal epithelial cell lines HT–29 was obtained from the American Type Culture Collection (ATCC, Rockville, MD). HT–29 cells were cultured in DMEM supplemented with 10% heat-inactivated fetal calf serum (FCS), 2 mM L-glutamine (Sigma), 1X Non essential Amino acid (Invitrogen), penicillin (50 IU/ml) and streptomycin (50 μg/ml) in an humidified atmosphere containing 10% CO2 at 37°C. After seeding, cells were grown 48h in 6 or 12 wells plate in antibiotic-free medium at 3.25X10⁵ and 6.5X10⁵ respectively. Medium was changed just before the addition of bacterial or reagents for 6h.
Rosiglitazone (used as positive control), GW9662, GW6471 and GW7647 (Cayman chemicals) were dissolved in DMSO following the manufacturer’s instructions and diluted at 100μM in antibiotic-free DMEM. They were used at a final concentration of 10μM except for GW6471 at 1μM. The antagonists (GW9662, GW6471) were added 1h before challenging with rosiglitazone or GW7647 respectively.

Jacouton E., Mach N., Cadiou J., Lapaque N., Clément K., Doré J., van Hylckama Vlieg J.E., Smokvina T, & Blottière H.M. (2015). Lactobacillus rhamnosus CNCMI-4317 Modulates Fiaf/Angptl4 in Intestinal Epithelial Cells and Circulating Level in Mice. PLoS ONE, 10(10), e0138880.

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THC Administration & PPAR Compound Prep

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Δ⁹-THC was provided by the NIDA pharmacy (Baltimore, MD). The stock solution was dissolved in ethanol at a concentration of 50 mg/ml. We diluted this solution as needed for experimental use in a 5% cremophor (Sigma-Aldrich, St. Louis, MO) saline solution. PPAR antagonists and agonists including GW9662, GW6471, pioglitazone, and GW7647 were purchased from Cayman Chemical (Ann Arbor, MI). Each compound was dissolved in a mixture of 2% DMSO, 3% tween-80 and 95% saline.

Hempel B., Crissman M., Pari S., Klein B., Bi G.H., Alton H, & Xi Z.X. (2023). PPARα and PPARγ are expressed in midbrain dopamine neurons and modulate dopamine- and cannabinoid-mediated behavior in mice. Molecular Psychiatry, 28(10), 4203-4214.

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Synthesis and Characterization of Phenylsulfonimide Derivatives

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The phenylsulfonimide derivatives IB42, IB44, and IB66 were synthesized, purified, and characterized as previously reported [25 (link)]. GW6471 was purchased from Cayman Chemical Company (Ann Arbor, MI, USA). Compounds were stored at −20 °C until the biological evaluation.

Gallorini M., Di Valerio V., Bruno I., Carradori S., Amoroso R., Cataldi A, & Ammazzalorso A. (2023). Phenylsulfonimide PPARα Antagonists Enhance Nrf2 Activation and Promote Oxidative Stress-Induced Apoptosis/Pyroptosis in MCF7 Breast Cancer Cells. International Journal of Molecular Sciences, 24(2), 1316.

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Muscle Metabolism Regulation in C57BL6 Mice

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C2C12 cells were obtained from Sigma (#91031101) along with Dulbecco's Modified Eagle Medium (DMEM; #D6429), fetal bovine serum (FBS; #F0926), horse serum (#H1270), and insulin (#I9278). Recombinant IL-15 was from GenScript (#Z03309-50) and GW-6471 (#11697) and GSK-3787 (#15219) were obtained from Cayman Chemicals. Trizol (#15596), SYBR green (#A25742) and a SuperScript VILO reverse transcription kit (#11754050) were purchased from ThermoFisher. The mitochondrial dye, Mito Red, was from Santa Cruz Biotechnology (SC-#301164). Male C57BL6 mice were kept on a 12 : 12 light dark cycle and fed a standard diet and water ad libitum. Gastrocnemius muscle was obtained following euthanasia. All animal procedures were approved by the Institutional Animal Care and Use Committee at Chapman University.

Thornton S.M., Krolopp J.E, & Abbott M.J. (2016). IL-15 Mediates Mitochondrial Activity through a PPARδ-Dependent-PPARα-Independent Mechanism in Skeletal Muscle Cells. PPAR Research, 2016, 5465804.

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NF-κB Transcriptional Activity Assay in A549 Cells

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A549 cells transfected with an NF-κB-dependent luciferase reporter gene were seeded in flat-bottomed 24-well plates at a concentration of 2.5 × 10⁴ cells per well. Cells were stimulated with 20 ng/mL tumor necrosis factor (TNF)-α (Peprotech, Rocky Hill, NJ, USA), following 24 h pre-incubation with test substances. Medium was removed 7 h after addition of TNF-α, and cells were lysed with 50 mM Tris-HCl buffer (pH 8.5) containing 1% Triton X-100. The transcriptional activity of NF-κB was determined by mixing cell lysates with a luciferase substrate (Steady-Glo Luciferase Assay system, Promega, Madison, WI, USA) according to the manufacturer’s instructions. Luminescence was measured using a Varioskan Flash 2.4 (Thermo Fisher Scientific Inc.). Similar experiments were conducted in the presence of a PPARα antagonist GW6471 (1 μM; Cayman Chemicals, Ann Arbor, MI, USA).

Nakanishi T., Motoba I., Anraku M., Suzuki R., Yamaguchi Y., Erickson L., Eto N., Sugamoto K., Matsushita Y, & Kawahara S. (2018). Naturally occurring 3RS, 7R, 11R-phytanic acid suppresses in vitro T-cell production of interferon-gamma. Lipids in Health and Disease, 17, 147.

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