All chemicals, dexamethasone and hydrocortisone (cortisol) were purchased from Sigma (St. Louis, MO), except Rosiglitazone (Enzo, Farmingdale, NY) and recombinant human insulin (Lilly, Indianapolis, IN). Collagenase type I was purchased from Worthington Biochemical (Lakewood, NJ). Cell culture media and fetal bovine serum (FBS) were obtained from Life Technologies (Carlsbard, CA). GR, MR and control siRNA were purchased from Qiagen and transfection reagents were purchased from Qiagen (HiPerFect, Germantown, MD) and Life Technologies (Lipofectamine and PLUS reagents, Carlsbard, CA).
Rosiglitazone
Manufactured by Enzo Life Sciences
Sourced in United States, Switzerland
Rosiglitazone is a laboratory research product used for scientific investigations. It is a synthetic compound that acts as a peroxisome proliferator-activated receptor gamma (PPARγ) agonist. The core function of Rosiglitazone is to modulate the activity of PPARγ, a nuclear receptor that plays a role in the regulation of glucose and lipid metabolism.
Automatically generated - may contain errors
Lab products found in correlation
Dexamethasone (dex), by Merck Group (6 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
D biotin, by Merck Group (3 mentions)
Collagenase type 1, by Worthington (2 mentions)
Lipofectamine and plus reagent, by Thermo Fisher Scientific (2 mentions)
Cell culture media, by Thermo Fisher Scientific (2 mentions)
Hiperfect, by Thermo Fisher Scientific (2 mentions)
Recombinant human insulin, by Enzo Life Sciences (2 mentions)
Hydrocortisone cortisol, by Merck Group (2 mentions)
Isobutylmethylxanthine, by Merck Group (2 mentions)
Humulin, by Eli Lilly (2 mentions)
Dmem f12, by Thermo Fisher Scientific (2 mentions)
Polybrene, by Merck Group (2 mentions)
Pantothenate, by Merck Group (2 mentions)
3 3 5 triiodo l thyronine sodium salt, by Merck Group (2 mentions)
Av 400, by Bruker (1 mentions)
Av 600, by Bruker (1 mentions)
3 isobutyl 1 methylxanthine ibmx, by Merck Group (1 mentions)
Antibiotic antimycotic, by Thermo Fisher Scientific (1 mentions)
D pantothenate, by Merck Group (1 mentions)
17 protocols using rosiglitazone
1
Glucocorticoid Receptor Signaling Assay
2
Characterizing Lipid Production and PPARγ in CN-G2 Cells
3
NMR and HPLC Analysis of Rosiglitazone
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The 1D and 2D NMR spectra were obtained with Bruker AV-400 or AV-600 instruments at 600\400 MHz for 1 H and 150\100 MHz for 13 C, CD3OD (δH 3.33; δ C 49.3). Prep-HPLC (Agilent 1260 Series) was performed on a C-18 column (SHISEID-PACK 20 mm × 250 mm, 5 um). The preparatory silica gels (100−300 mesh) and Sephadex LH-20 were obtained from QMC Co., Ltd. and GE-H Co., Ltd.31 (link).
Rosiglitazone was purchased from Enzo Life Science (Lausen, Switzerland). High-fat diet (HFD, Product# D12492) and low-fat diet (LFD Product# D12450B) were obtained from Research Diets (New Brunswick, NJ, USA). HFD with 60 kcal% fat includes 26.2% protein, 26.3% carbohydrate, and 34.9% fat. LFD with 10 kcal% fat includes 19.2% protein, 67.3% carbohydrate and 4.3% fat.
Rosiglitazone was purchased from Enzo Life Science (Lausen, Switzerland). High-fat diet (HFD, Product# D12492) and low-fat diet (LFD Product# D12450B) were obtained from Research Diets (New Brunswick, NJ, USA). HFD with 60 kcal% fat includes 26.2% protein, 26.3% carbohydrate, and 34.9% fat. LFD with 10 kcal% fat includes 19.2% protein, 67.3% carbohydrate and 4.3% fat.
4
Glucocorticoid Receptor Signaling Assay
5
Adipogenic Differentiation of hMADS Cells
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Human multipotent abdominal subcutaneous adipose-derived mesenchymal stem cells (hMADS) were obtained from overweight/obese men with impaired glucose metabolisms and pooled and differentiated into the adipogenic lineage. Therefore, cells were seeded at a density of 2000 cells∙cm−2 and cultured, as described previously [27 ,28 (link)]. Human multipotent adipose-derived mesenchymal stem cells (hMADS) were kept in a proliferation medium containing DMEM and Ham's F-12 (DMEM-Ham's F-12) Nutrient Mixture (no. 31330-095, Gibco), 10 % FBS (Bodinco BV), and 1x Antibiotic-Antimycotic (Gibco). At ~80 % confluence, a differentiation medium was added to the cells containing DMEM-Ham's F-12, 3 % FBS (Bodinco BV), 1x Antibiotic-Antimycotic (Gibco), 33 μM D-Biotin (no. B4693, Sigma), 17 μM d -pantothenate (no. P5155, Sigma), 0.1 μM h-insulin (no. 91077C, Sigma), 1 μM dexamethasone (no. D4902, Sigma), 250 μM 3-isobutyl-1-methylxanthine (IBMX, no. I5879, Sigma), and 5 μM rosiglitazone (no. ALX-350-125-M025, Enzo Life Sciences). After 7 days, IBMX and rosiglitazone were removed from the medium. All cells were proliferated and differentiated under 21 % O2, and thereafter, exposed to either 10 % O2 continuously (resembling physiological normoxia), or to MIH consisting of 3 × 2 h cycles per day, alternating between 5 and 10 % O2, during the final 7 days of cell culture.
6
Differentiation of 3T3-L1 Fibroblasts to Adipocytes
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3T3-L1 mouse fibroblasts were cultured at confluence and induced to differentiate in mature adipocytes by two differentiation mixes. Thus, control (i.e., DMEM-5% FBS or DMEM-5% FBS plus LPS 0.5 ng/mL) or conditioned media (collected from LPS stimulating and not J774A.1 macrophages) were supplemented by isobutyl methylxanthine 0.5 µM (Sigma-Aldrich Inc., St. Louis, MO, USA, Cat# I5879), dexamethasone 0.25 µM (Sigma-Aldrich Inc., St. Louis, MO, USA, Cat# D1756), rosiglitazone 10 µM (Enzo Life Science, Farmingdale, NY, USA, Cat# ALX-350-125) and insulin 5 µg/mL (Humulin, Lilly, Indianapolis, IN, USA, Cat# HI0290). After two days, the cells were treated with insulin (5 µg/mL) and rosiglitazone (10 µM) for an additional two days. Then, fresh medium—i.e., without the adipogenic cocktail—was added until the adipocyte phenotype appeared in more than 90% of the cells (i.e., 8 days after induction) by microscopic visualization.
7
CCl4-Induced Hepatic Fibrosis in Mice
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Fibrosis was induced in male C57BL/6 mice (20-24 g, Charles River Laboratories, Wilmington, MA) as described[15 (link)]. Briefly, mice received twice-weekly intraperitoneal injections of CCl4 (diluted 1:7 in sunflower oil) at 1 mL/kg body weight, for a total of 6 wk to induce hepatic fibrosis. Control (nonfibrotic) mice received oil alone. For the final 2 wk of treatment, mice were randomly assigned to receive implantation of subcutaneous osmotic pumps (model 1002, Durect, Cupertino CA) to deliver serelaxin (generously provided by Dennis Stewart, Novartis) at 150 μg/g per day, or vehicle (citrate buffer). Rosiglitazone (4 mg/kg per day Enzo Life Sciences, Farmingdale, CA) or vehicle (5% DMSO in phosphate buffered saline) was also administered daily by oral gavage for the final 2 wk. Each group contained 5 mice. Mice were sacrificed 72 h after the final CCl4 injection, and liver and blood were collected. Mice were maintained at 22 °C under 12-h light/dark cycles, and had free access to food and water throughout the study. All procedures were conducted in accordance with The Guide for the Care and Use of Laboratory Animals[27 ], and were approved by the VA Nebraska Western-Iowa Institutional Animal Care and Use Committee.
8
Adipogenic Differentiation Assay Protocol
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Cells were plated at a density of 5 × 104 per well in 24-well plates. After overnight incubation, adipogenic induction medium with 5 μg/ml insulin (SIGMA), 1 μM Dexamethason (SIGMA), 500 μM 3-isobutyl-1-methylxanthine (IBMX), 1 μg/ml rosiglitazone (ENZO Lifesciences) in DMEM-F12 was applied for 2 days. Subsequently, cells were cultured for another 2 days in DMEM-F12 with 5 μg/ml insulin, 10% FBS, 1% streptomycin/penicillin and for 2 more days in the medium without insulin. For Oil Red O staining, cells were rinsed twice with PBS and fixed with 10% formalin in PBS for 5 min at RT. Cells were subsequently fixed for 1 h at RT with fresh formalin solution, washed with 60% isopropanol and then completely dried. Oil Red O working solution, made up of 6 parts filtered Oil Red O stock (0.7 g Oil Red O in 200 ml isopropanol; Sigma) and 4 parts distilled water, was applied to the dried well for 10 min incubation. Cells were rinsed with distilled water 4 times and dried prior to microscopic observation.
9
Lentiviral Infection and Adipogenic Differentiation of Human Myoblasts
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Human myoblasts were grown in the incomplete SkBM medium until 90-100% confluence in 12-well plates and were infected by exposure to lentiviral supernatant (∼ 4 MOI) and polybrene (0.8 μg/mL, Millipore, Billerica, MA) daily for two days. After the second lentiviral infection, human myoblasts or AdSVCs underwent adipogenesis according to the non-modified adipogenesis protocol (23 (link)). Briefly, the cells were cultured in the adipocyte induction medium DMEM/F12 (Gibco) containing d-Biotin (33 nM, Sigma), human insulin (70 nM, Sigma I9278), dexamethasone (100 nM, APP pharmaceuticals, Schaumburg, IL), pantothenate (4 μg/mL, Sigma), human transferrin (10 μg/mL, Calbiochem), 3,3′, 5-triiodo-L-thyronine sodium salt (2 μM, Sigma), rosiglitazone (1μM, Enzo Lifesciences, Farmingdale, NY), isobutylmethylxanthine (IBMX, 0.5 mM, Sigma), 10% FBS, 100U/mL penicillin-streptomycin. After 72 hr, cells were switched to the adipocyte differentiation medium (induction medium without rosiglitazone and IBMX) for additional 8 days or until collection.
10
Preparation and Storage of Bioactive Compounds
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Simvastatin and vorinostat purchased from Cayman Chemical (Ann Arbor, MI, USA), romidepsin, belinostat and entinostat purchased from Selleck Chemicals (Houston, TX, USA), panobinostat purchased from LC Laboratories (Boston, MA, USA), and rosiglitazone and tunicamycin purchased from Enzo Life Sciences (Farmingdale, NY, USA) were dissolved in dimethyl sulfoxide (DMSO). Compound C dihydrochloride purchased from R&D Systems (Minneapolis, MN, USA) and cycloheximide (CHX) purchased from Enzo Life Sciences were dissolved in distilled water. These reagents were stored at −80 °C or −20 °C until use.
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