Antibodies against c-Src, phospho-Src (Tyr416), NF-κB p65, phospho-NF-κB p65 (Ser536), COX-2, PPARγ, and UCP2 were purchased from Cell Signaling Technology (Beverly, MA). Antibodies against fibronectin and collagen IV were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Antibodies against E-cadherin, α-SMA, DRP1, and FIS1 were purchased from the Proteintech Group (Chicago, USA). The β-actin antibody was purchased from Abcam (Cambridge, UK). The goat anti-rabbit IgG-HRP secondary antibody was purchased from Zhongshan Golden bridge Biotechnology (Beijing, China). c-Src inhibitor PP2, NF-κB inhibitor PDTC, COX-2 inhibitor NS-398, and PPARγ antagonist GW9662 were purchased from Selleck Chemicals (Houston, TX, USA). The pIRES2-ZsGreen1-UCP2 overexpression plasmid and PLV-UCP2-shRNA plasmid were purchased from Yingrun Biotechnology Inc. (Changsha, China). PhosSTOP phosphatase inhibitor cocktail tablets were purchased from Roche Ltd. (Mannheim, Germany). The protease inhibitor cocktail was purchased from Calbiochem (San Diego, CA, USA). All culture media were purchased from Gibco-BRL (Grand Island, NY, USA). D-glucose was purchased from Sigma (St. Louis, MO, USA). The polyvinylidene difluoride (PVDF) membrane was purchased from Millipore (Billerica, MA, USA).
Gw9662
Manufactured by Selleck Chemicals
Sourced in United States, China
GW9662 is a synthetic organic compound that functions as a selective antagonist of the peroxisome proliferator-activated receptor gamma (PPARγ). It is commonly used in scientific research applications.
Automatically generated - may contain errors
Lab products found in correlation
Rosiglitazone, by Selleck Chemicals (4 mentions)
T0070907, by Selleck Chemicals (4 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
Foetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Dulbecco s modified eagle s medium, by Thermo Fisher Scientific (2 mentions)
Trypsin edta, by Thermo Fisher Scientific (2 mentions)
Dexamethasone (dex), by Merck Group (2 mentions)
Insulin, by Merck Group (2 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (2 mentions)
D glucose, by Merck Group (1 mentions)
β actin antibody, by Abcam (1 mentions)
Phosstop phosphatase inhibitor cocktail tablet, by Roche (1 mentions)
Polyvinylidene difluoride pvdf membrane, by Merck Group (1 mentions)
Pparγ, by Cell Signaling Technology (1 mentions)
Recombinant human il 33, by R&D Systems (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Beyotime (1 mentions)
Cycloheximide (chx), by Selleck Chemicals (1 mentions)
Bafilomycin a1, by Selleck Chemicals (1 mentions)
Rapamycin, by Selleck Chemicals (1 mentions)
Il 33, by R&D Systems (1 mentions)
25 protocols using gw9662
1
Molecular Mechanism of UCP2 Regulation
2
Comprehensive Autophagy and Signaling Pathway Analysis
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The antibodies used in our experiments were: LC3 (CST, #2775), IL-33 (R & D, AF3626 and AF4810), ST2 (Thermofisher, PA5-20077), p-STAT3 (CST, #9145), STAT3 (CST, #9139), p-AMPK (CST, #2535), AMPK (CST, #2532), p-ULK1 (Thermofisher, PA5-105129), ULK1 (Abclonal, A8529), p62 (Abclonal, A1970), GKN-1 (Proteintech, 19344-1-AP), GKN-2 (Abcam, ab188866), GKN-3 (Cloud-clone, PAK528Mu01), Tubulin (Proteintech, 66031-1-Ig).
The chemicals used in our experiments were: Bafilomycin A1 (Selleck, S1413), 3-MA (Selleck, S2767), Rapamycin (Selleck, S1039), MNNG (Selleck, E0157), NAC (Selleck, S1623), Cycloheximide (Selleck, S7418), DAPI (Beyotime, C1002), Recombinant Human IL-33 (R & D, 3625-IL), Pifithrin-α HBr (Selleck, E0157), 2-MeOE2 (Selleck, S1233), ML385 (Selleck, S8790), SR11302 (MCE, HY-15870), BAY 11 (Selleck, S2913), GW9662 (Selleck, S2915), IQ3 (Selleck, S0781), STAT3-IN-7 (Selleck, S0986), FH535 (Selleck, S7484).
The chemicals used in our experiments were: Bafilomycin A1 (Selleck, S1413), 3-MA (Selleck, S2767), Rapamycin (Selleck, S1039), MNNG (Selleck, E0157), NAC (Selleck, S1623), Cycloheximide (Selleck, S7418), DAPI (Beyotime, C1002), Recombinant Human IL-33 (R & D, 3625-IL), Pifithrin-α HBr (Selleck, E0157), 2-MeOE2 (Selleck, S1233), ML385 (Selleck, S8790), SR11302 (MCE, HY-15870), BAY 11 (Selleck, S2913), GW9662 (Selleck, S2915), IQ3 (Selleck, S0781), STAT3-IN-7 (Selleck, S0986), FH535 (Selleck, S7484).
3
Modulation of GPER1, ERs, and Signaling
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The ICP-1 cells were seeded in 6-well plates and pre-incubated with 1 μM GPER1 inhibitor G15 (Cat#S6651, Selleck, Houston, TX) for 12 h, 1 μM GPER1 activator G1 (Cat#S0851, Selleck) for 1 h, 1 μM estrogen receptor antagonist Fulvestrant (Cat#S1191, Selleck) for 12 h, 10 μM ERβ antagonist PHTPP (Cat#S8686, Selleck) for 1 h, 10 μM Erk inhibitor U0126 (Cat#S1102, Selleck) for 4 h or 5 μM PPARγ inhibitor GW9662 (Cat#S2915, Selleck) for 4 h, respectively. Then, the cells were treated with 0 or 40 μM GEN for another 24 h. After that, the protein levels of ERα, ERβ, GPER1, p-Erk, t-Erk and PPARγ were assessed using Western blot.
4
Wnt/β-catenin and PPARγ Regulation of Osteogenesis
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Foetal bovine serum (FBS) and Dulbecco's Modified Eagle's medium (DMEM) were obtained from Gibco‐BRL (Thermo Fisher Scientific, Inc). β‐Glycerophosphate (β‐GP; G5422) and adenine (V900471) were purchased from Sigma‐Aldrich (Merck KGaA). Ginsenoside Rb1 (41753‐43‐9) was obtained from Fleton Natural Products Co., Ltd. The Wnt/β‐catenin agonist SKL2001 (S8320) and PPAR‐γ inhibitor GW9662 (S2915) were from Selleck Chemicals. The Nuclear and Cytoplasmic Protein Extraction Kit (P0027) was from Beyotime Biotechnology.
Antibodies against calponin 1 (#17819), runt‐related transcription factor 2 (RUNX2; #12556), β‐catenin (#8480), phospho‐β‐catenin (Ser675) (#9567), glycogen synthase kinase‐3β (GSK‐3β; #9315), phospho‐GSK‐3β (Ser9) (#9322) and histone‐H3 (#4499) were from Cell Signaling Technology, Inc. An antibody against α‐smooth muscle actin (α‐SMA; NBP2‐22120) was purchased from Novus Biologicals. An antibody against peroxisome proliferator‐activated receptor gamma (PPAR‐γ; sc‐7273) was purchased from Santa Cruz Biotechnology. An antibody against glyceraldehyde‐3‐phosphate dehydrogenase (GAPDH; TA‐08) was purchased from ZSGB‐BIO.
Antibodies against calponin 1 (#17819), runt‐related transcription factor 2 (RUNX2; #12556), β‐catenin (#8480), phospho‐β‐catenin (Ser675) (#9567), glycogen synthase kinase‐3β (GSK‐3β; #9315), phospho‐GSK‐3β (Ser9) (#9322) and histone‐H3 (#4499) were from Cell Signaling Technology, Inc. An antibody against α‐smooth muscle actin (α‐SMA; NBP2‐22120) was purchased from Novus Biologicals. An antibody against peroxisome proliferator‐activated receptor gamma (PPAR‐γ; sc‐7273) was purchased from Santa Cruz Biotechnology. An antibody against glyceraldehyde‐3‐phosphate dehydrogenase (GAPDH; TA‐08) was purchased from ZSGB‐BIO.
5
Modeling Liver Injury in Mice
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C57BL/6 mice were purchased from the Shanghai Laboratory Animal Center of the Chinese Academy of Sciences. 129Sv/Ev wild-type (WT) mice were obtained from the Shanghai Xipuer-Bikai Laboratory Animal Limited Company. Bex1-deficient (Bex1−/−) mice were kindly provided by Professor Frank L. Margolis, University of Maryland, Baltimore, MD, USA [21 (link)]. Female mice 6–8 weeks old were administered a CDE diet (TROPHIC, Nantong, China) supplemented with 0.15% (w/v) d ,l -ethionine (Sigma-Aldrich, St. Louis, MO, USA) in the drinking water for 3 weeks [23 (link)], while control mice received normal chow and drinking water. Rosiglitazone (TCI, Tokyo, Japan) or GW9662 (dose of 2 mg/kg; Selleckchem, Houston, TX, USA) was administered to mice via intraperitoneal injection every second day, for a dose of 50 mg/kg, and dimethyl sulphoxide (DMSO; Sigma-Aldrich) was injected into control mice. All mice were maintained under specific pathogen-free conditions in the vivarium of Shanghai Jiao Tong University School of Medicine. All animal procedures were approved by the Animal Welfare & Ethics Committee of Shanghai Jiao Tong University School of Medicine.
6
Evaluating Fibrosis Regulators in Cell Culture
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The phorbol 12-myristate 13-acetate (PMA) was purchased from CSNpharm (China, CSN11701) and dissolved in DMSO. The LPS (Sigma-Aldrich, USA, L2630), TGF-β1 (SinoBiological, China, 10804-HNAC) and CCL2 (GenScript, China, Z03292) were dissolved in H2O. The Z-guggulsterone/GS (Selleck, Houston, S6812), GW9662 (Selleck, Houston, S2915), GW4064, Rosiglitazone/RSG and GW3965 (MCE, China, HY-50108, HY-17386 and HY-10627A) were dissolved in DMSO. The antibodies used were as follows: Collagen I and p-Smad3 (abcam, USA, ab260043, ab52903), t-Smad3 and GAPDH (Cell Signaling Technology, USA, 9523, 5174), α-SMA (Proteintech, 23081-1-AP).
7
Evaluating Kuhuang Injection's Hepatoprotective Effects
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DMEM was purchased from Thermo Fisher Biochemicals Co., Ltd. (USA). Foetal bovine serum, PLA2 enzyme, and penicillin were purchased from Gibco (USA). An AMV one-step RT-PCR kit was obtained from TaKaRa (China). Kuhuang injection extract powder was provided by Changshu Leiyunshang Pharmaceutical Co., Ltd. (China). A peristaltic pump was purchased from Master-Flex (German), and a 0.22 μm filter (Millex-GV) (German) was used. Sodium Deoxycholate (SDC) was obtained from VETEC (USA), and disposable indwelling intravenous needles (BD type Y 22 G) (USA) were used. A cell counting kit-8 (CCK-8) kit was purchased from Dojindo (USA), and reactive oxygen species (ROS) detection kits were purchased from Shanghai Biyuntian (China). Malondialdehyde (MDA) assay kits, superoxide dismutase (SOD) assay kits, glutathione (GSH) assay kits, and TBA assay kits were purchased from Nanjing Jiancheng Institute of Bioengineering (China). Aspartate transaminase (AST) test kits, alanine transaminase (ALT) test kits, lactate dehydrogenase (LDH) test kits, and alkaline phosphatase (ALP) test kits were all obtained from Nanjing Jiancheng Institute of Biological Engineering (China). CPZ was purchased from MCE (MedChemExpress) (USA), and GW9662 was obtained from Selleck (USA).
8
Cell Viability Assay with Rosiglitazone and GW9662
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The cells were placed in 96-well plates at a density of 5 × 103 cells/well and cultured overnight. Then, the cells were cultured with a complete medium containing different concentrations of rosiglitazone (R2408, Sigma-Aldrich; Merck KGaA) or GW9662 (No.S2915, Selleck Chemicals, Houston, TX, USA) for 24, 48, and 72 h. After 24, 48 and 72 h, cells were incubated with CCK8 solution (10 μl/well, Beyotime Biotechnology, no. C0038) for 1 h at 37 °C. The absorbance was measured at OD 450 nm by a microplate reader (Thermo Fisher Scientific Inc.).
9
Therapeutic Compounds for Post-SAH Treatment
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Drug preparation and administration were previously described by Zhong et al.28 (link) Briefly, 20 mg bexarotene (Selleck, Shanghai, China) was dissolved in 5.7 ml
dimethyl sulfoxide (DMSO), and 51.3 ml phosphate-buffered saline (PBS) was added to the
mixture. Similarly, 10 mg GW9662 (Selleck), an inhibitor of PPARγ was dissolved in 3.6 ml
DMSO, and PBS (32.4 ml) was added to the mixture. A total of 10 mg of MK886 (Selleck), an
antagonist of FLAP was dissolved in 2.1 ml DMSO, and 18.9 ml PBS was added to the mixture.
The final concentrations of GW9662, MK886 and bexarotene were the same (1 mM). The
solution was injected intraperitoneally at 5 mg/kg bexarotene, 2 mg/kg GW9662, and 3 mg/kg
MK886 per day after SAH. The vehicle group solution was prepared in the same manner and
administered in the same volume, same route and same time course as above. These reagents
were used immediately after SAH and continued to be used once a day until the rats were
euthanized.
dimethyl sulfoxide (DMSO), and 51.3 ml phosphate-buffered saline (PBS) was added to the
mixture. Similarly, 10 mg GW9662 (Selleck), an inhibitor of PPARγ was dissolved in 3.6 ml
DMSO, and PBS (32.4 ml) was added to the mixture. A total of 10 mg of MK886 (Selleck), an
antagonist of FLAP was dissolved in 2.1 ml DMSO, and 18.9 ml PBS was added to the mixture.
The final concentrations of GW9662, MK886 and bexarotene were the same (1 mM). The
solution was injected intraperitoneally at 5 mg/kg bexarotene, 2 mg/kg GW9662, and 3 mg/kg
MK886 per day after SAH. The vehicle group solution was prepared in the same manner and
administered in the same volume, same route and same time course as above. These reagents
were used immediately after SAH and continued to be used once a day until the rats were
euthanized.
10
Antibody-based Protein Signaling Assay
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Primary antibodies detecting PPARγ, Phosphatase and tensin homolog (PTEN), phospho-Akt (Ser473), p21 Waf1/Cip1, phospho-Bad (Ser136), Phospho-FoxO1 (Ser256), Phospho-Bcl2 (Ser70), and HRP-labeled secondary antibodies were purchased from Cell Signaling Technology (MA, USA). Antibodies detecting phospho-Akt (Ser473) and PPARγ in the immunohistochemistry were purchased from LifeSpan (Seattle, WA, USA). The PPARγagonists rosiglitazone and pioglitazone, the inverse-agonist T0070907, and the antagonist GW9662 were purchased from Selleck Chemicals (TX, USA).
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