For CML (MyBioSource, San Diego, USA, Catalogue No.: MBS390113) stimulating, cells (5×103) in 100 μl DMEM were added to 96-well plates with different concentrations of CML (0, 12.5, 25, 50, 75, and 100μg/ml) for 6, 12 and 24 hours. GLP-1 (Bioss, Beijing, China, Catalogue No.: bs-0038P) treatment was performed with GLP-1 at concentrations of 0nM, 50nM, 100nM, 200nM, and 500nM in the medium with cells stimulated by 50 μg/ml CML for 24 hours. GW9662 (MedChemExpress LLC, Shanghai, China, Catalogue No.: 22978-25-2) blocking administration was conducted by 0, 1, 5, 10, 20, and 50 μM GW9662 in PC12 cells with 50 μg/ml CML and 100nM GLP-1 for 24 hours. After CML stimulating, GLP-1 treatment and GW9662 blocking assay, CCK-8 assay was performed according to the protocol of manufacturer (Jiangsu KeyGEN BioTECH Corp., Ltd, Nanjing, China, Catalogue No.: KGA317-2).
Gw9662
Manufactured by MedChemExpress
Sourced in United States, China
GW9662 is a lab equipment product manufactured by MedChemExpress. It functions as a high-affinity antagonist for the peroxisome proliferator-activated receptor gamma (PPARγ).
Automatically generated - may contain errors
Lab products found in correlation
Rosiglitazone, by MedChemExpress (6 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (4 mentions)
Reverse transcription kit, by Thermo Fisher Scientific (3 mentions)
Imipramine hydrochloride imi, by Merck Group (2 mentions)
Lipofectamine 2000 reagent, by Thermo Fisher Scientific (2 mentions)
Dmem f12 medium, by Keygen Biotech (2 mentions)
Paraformaldehyde (pfa), by Merck Group (2 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (2 mentions)
Trizol reagent, by Thermo Fisher Scientific (2 mentions)
Foetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Propofol, by Merck Group (2 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions)
Sodium valproate, by Merck Group (1 mentions)
Oil red o powder, by Merck Group (1 mentions)
Milli q water purification system, by Merck Group (1 mentions)
Ammonium acetate, by Merck Group (1 mentions)
Ly294002, by MedChemExpress (1 mentions)
Agilent tc c18, by Agilent Technologies (1 mentions)
Huvecs, by Merck Group (1 mentions)
34 protocols using gw9662
1
Evaluating CML-Induced Cell Viability
2
Lipid Extraction and Analysis
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Liquid-chromatography-grade acetonitrile, methanol, dichloromethane, and isopropanol were purchased from Fisher Scientific (Pittsburgh, PA, USA). Internal standard TAG 45:0, sodium valproate, Oil Red O powders, and ammonium acetate were purchased from Sigma-Aldrich (St. Louis, MO, USA). GW9662, rosiglitazone, and LY294002 were purchased from MedChemExpress (Shanghai, China). Ultrapure water was prepared using a Milli-Q water purification system procured from Millipore Corp. (Billerica, MA, USA).
3
Pharmacokinetics of Ginsenoside Rb1 in Hippocampus
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GRb1 (purity ≥ 97%, C54H92O23, Cat# P0088) was purchased from the Pureone Biotechnology Company of Shanghai. Lipopolysaccharide (LPS, E. coli, 0127: B8) were purchased from Sigma-Aldrich Chemical Co. (St Louis, MO, USA) and soluble in phosphate-buffered saline (PBS, Servicebio, Cat# G4202). The experiment administered the following treatments once per day for 4 weeks (at 16:00 h): Saline, GRb1 (20 mg/kg/d, given as a 2 mg/ml solution in 0.9% saline, intragastric administration; Herbpurify, Chengdu, China), or imipramine hydrochloride (IMI, 20 mg/kg/d, intraperitoneally (i.p.); Sigma-Aldrich, Darmstadt, Germany). A subset of stressed animals was pretreated with PPARγ inhibitor GW9662 (1 mg/kg/day in 1% DMSO, i.p., 28 d; Med Chem Express, Monmouth Junction, NJ, USA), then treated 1 h later with GRb1 (control mice were administrated with 1% DMSO solution in 0.9% saline). The doses of GRb1 and IMI were chosen based on previous studies [30 (link), 31 (link)]. After intragastric administration for 0.5 h, 1 h, 2 h, 3 h, 12 h, 24 h, and 48 h, the concentration of GRb1 in hippocampus tissues was detected by LC-MS/MS technique.
4
HPLC Analysis of DGLHD Quality
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Quality control of DGLHD was analyzed by HPLC on a reverse-phase analytical column (Agilent TC-C18, 4.6 × 250 mm, Agilent Technologies). The mobile phase was made up of (A) 100% water with 0.1% phosphoric acid and (B) 100% acetonitrile. Gradient elution from 5 to 45% B for 40 min was used at 10 min intervals. The flow rate was 1 mL/min and the detection wavelength was 280 nm. Chemical standards included calycosin-7-glucoside, verbascoside, ferulic acid, coptisine, berberine, baicalin, wogonoside, baicalein, and wogonin purchased from the National Institutes of Food and Drug Control (Beijing, China). Their purity was verified by HPLC. And GW9662 (PPAR-γ antagonist) was purchased from MedChem Express (Shanghai, China).
5
Establishing Atherosclerosis Cell Models
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Human umbilical vein endothelial cells (HUVECs) were purchased from Sigma-Aldrich (St. Louis, MO, USA) and maintained in DMEM (Gibco; Thermo Fisher Scientific, Inc.) with 10% fetal bovine serum (FBS, Gibco) and 1% penicillin–streptomycin (Gibco) at 37°C. Medium was refreshed every other day unless stated otherwise. HUVECs were incubated 40 μg/mL of ox-LDL (Guangzhou Yiyuan Biotechnologies, Guangzhou, China) for 24 h to establish atherosclerosis cell models in vitro. A PPAR-γ antagonist GW9662 (MedChemExpress, Shanghai, China) was used to disturb PPAR-γ signaling.
6
Ox-LDL and DiI-Ox-LDL Signaling Pathway
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Ox-LDL and DiI-Ox-LDL were purchased from Yiyuan Biotechnology (Guangzhou, China). Antibodies against PPAR-γ (57 kDa, ab59256), LXR-α (50 kDa, ab41902), and ABCA1 (254 kDa, ab18180) were purchased from Abcam (Cambridge, MA, USA). β-Actin (42 kDa, #3700) was purchased from Cell Signaling Technology (Beverly, MA, USA). GW9662 was obtained from MedChemExpress (Monmouth Junction, NJ, USA).
7
Macrophage Activation by SARS-CoV-2 and R848
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Primary human macrophages were obtained by plating fresh or frozen splenocytes obtained from donors with ruptured spleen caused by trauma with RPMI 1640 medium with 50 ng/mL M-CSF (Novoprotein, Suzhou, Jiangsu, China) and removing unattached cells after 12 hours of culturing. The attached macrophages were then cultured in complete RPMI-1640 medium or RPMI-1640 medium without glutamine and pyruvate (Seahorse Bioscience, Santa Clara, California, USA) for 12 h before simulation with R848 (Sigma, St. Louis, Missouri, USA) or SARS-CoV-2 (MOI = 1). In other experiments, macrophages were treated with 2-DG (1mM; Sigma, St. Louis, Missouri, USA), PPARγ inhibitor GW9662 (2μM; MedChemExpress, Monmouth Junction, NJ, USA), or PPARγ activator rosiglitazone (5μM; MedChemExpress, Monmouth Junction, NJ, USA) for 48 h before stimulation with R848 or SARS-CoV-2 as indicated (MOI = 1). After 24 h of stimulation, supernatants were collected for the measurement of IL-6, TNF-α, CCL2, CCL3 by ELISA and cells were harvested for the detection of IL1B, CD36, and FABP4 mRNA by real-time PCR.
8
Investigating Pharmacological Modulators
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3-NP, dimethyl sulfoxide (DMSO), and carboxymethyl cellulose (CMC) were procured from Sigma-Aldrich, St. Louis, MD, USA. Telmisartan (Micardis®) was purchased from Boehringer Ingelheim, Germany. Pioglitazone (Actos®) was purchased from Takeda Pharmaceuticals Ltd. The PPARγ antagonist, GW9662, was purchased from MedChemExpress, NJ, USA. All other chemicals and reagents used in this study were of analytical grade.
9
Investigating the Role of Nrf2 in NOD Mice
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Non-obese diabetic (NOD) mice and Nrf2−/− mice were obtained from the Model Animal Research Center of Nanjing University (Nanjing, China). C57BL/6 mice were from Comparative Medical Center of Yangzhou University (Yangzhou, China). Experiments used 6–12-week-old and sex-matched mice, unless otherwise indicated. All animal care and handling procedures were conducted in accordance with the protocols approved by the Institutional Animal Care and Use Committee (IACUC) at Yangzhou University (Yangzhou, China).
The following reagents were used: Astilbin (HY-N0509); 1-aminobenzotriazole (ABT, HY-103389), a nonspecific and irreversible inhibitor of CYP enzymes; etomoxir (HY-50202), an inhibitor of CPT1α; SF1670 (HY-15842), an inhibitor of PTEN; N-Acetyl-L-cysteine (NAC, HY-B0215), a GSH-related antioxidant; GW9662 (HY-16578), a potent and selective PPARγ antagonist; and dorsomorphin (Compound C, HY-13418A), a selective and ATP-competitive AMPK inhibitor (MedChemExpress, NJ, United States).
The following reagents were used: Astilbin (HY-N0509); 1-aminobenzotriazole (ABT, HY-103389), a nonspecific and irreversible inhibitor of CYP enzymes; etomoxir (HY-50202), an inhibitor of CPT1α; SF1670 (HY-15842), an inhibitor of PTEN; N-Acetyl-L-cysteine (NAC, HY-B0215), a GSH-related antioxidant; GW9662 (HY-16578), a potent and selective PPARγ antagonist; and dorsomorphin (Compound C, HY-13418A), a selective and ATP-competitive AMPK inhibitor (MedChemExpress, NJ, United States).
10
Modulation of LCWE-Induced Inflammation in Mice
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In total 20, male (wild-type) C57BL/6 mice (age, 4–6 weeks; weight, 18–20 g) were obtained from the Animal Centre of Shandong Medical University (Shandong, China). All mice were maintained under specific pathogen-free conditions (20–26°C, 40–70% humidity, 12-h light/dark cycle with access to full-valence granular rat feedstuff and sterile water ad libitum). Mice were randomly divided into four groups: PBS, LCWE, LCWE+AUDA and LCWE+AUDA+GW9662. In each group, mice were injected intraperitoneally with 0.5 ml PBS alone; PBS supplemented with 0.5 mg LCWE; PBS supplemented with 0.5 mg LCWE and 10 mg/kg AUDA (Cayman Chemical, Wuhan, China); or PBS supplemented with 0.5 mg LCWE, 10 mg/kg AUDA and 10 mg/kg GW9662 (MedchemExpress, Shanghai, China), respectively. Following 14-day induction, mice were sacrificed.
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