Cell cultures were maintained as previously described[85 (link)]. HEK-293T and HepG2 cells were maintained in Dulbecco’s modified Eagle’s medium supplemented with 10% fetal bovine serum and 1% antibiotic/antimycotic (Thermo Fisher) at 37°C, 5% CO2. A549 (ATCC) cells were maintained in Ham’s F-12K (Kaighn’s) medium supplemented with 10% fetal bovine serum and 1% antibiotic/antimycotic at 37°C, 5% CO2. H460 (ATCC) cells were maintained in RMPI-1640 medium supplemented with 10% fetal bovine serum and 1% antibiotic/antimycotic at 37°C, 5% CO2. Transfections were performed with Lipofectamine 3000 (Thermo Fisher) according to the manufacturer’s protocol. siRNAs (Thermo Fisher) used include: KEAP1 (HSS114799, HSS114800, HSS190639), WDR23 (HSS129631, HSS129632, HSS129633), NRF2 (s9492). For genes with more than one siRNA listed, a cocktail mixture of the previously mentioned siRNAs is used for efficient knockdown. Chemical treatments include: 50μM tert-Butylhydroquinone (Sigma) and 250μM H2O2 (Sigma). 50μM tert-Butylhydroquinone (Sigma), 250μM H2O2 (Sigma), and 10μM MG-132 (Sigma).
Tert butylhydroquinone
Manufactured by Merck Group
Sourced in United States, Germany
Tert-butylhydroquinone is a chemical compound used as a preservative and antioxidant in various industrial applications. It is a white crystalline solid with the chemical formula C₁₁H₁₆O₂.
Automatically generated - may contain errors
Lab products found in correlation
Gallic acid, by Merck Group (3 mentions)
Tert butyl hydroperoxide, by Merck Group (2 mentions)
Curcumin, by Merck Group (2 mentions)
Sulforaphane, by Merck Group (2 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Mg132, by Cayman Chemical (2 mentions)
Folin ciocalteu reagent, by Merck Group (2 mentions)
2 2 diphenyl 1 picrylhydrazyl (dpph), by Merck Group (2 mentions)
Acetic acid, by Merck Group (2 mentions)
Lipofectamine 3000, by Thermo Fisher Scientific (1 mentions)
Mg132, by Merck Group (1 mentions)
Antibiotic antimycotic, by Thermo Fisher Scientific (1 mentions)
Seleno l methionine, by Merck Group (1 mentions)
Nec 1, by Merck Group (1 mentions)
N acetylcysteine, by Merck Group (1 mentions)
Q vd oph, by Merck Group (1 mentions)
Dimethyl fumarate, by Merck Group (1 mentions)
Dimethyl itaconate, by Merck Group (1 mentions)
Trans chalcone, by Merck Group (1 mentions)
25 protocols using tert butylhydroquinone
1
Cell Culture Maintenance and Transfection Protocols
2
Antioxidant Agents for Cell Assays
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Seleno-L-methionine (SeMet; ≥98% purity), tert-Butyl hydroquinone (tBHQ; 97% purity), and tert-Butyl hydroperoxide (tBOOH; 70% solution in water) were purchased from Sigma-Aldrich (Oakville, ON, Canada).
3
Compound Sources for Biological Assays
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The compounds dimethyl fumarate (DMF), trans-chalcone, curcumin, sulforaphane, tert-butylhydroquinone (tBHQ) and dimethyl itaconate (DMI) were purchased from Sigma-Aldrich. 4-OI was chemically synthetized as described elsewhere [9 (link)]. The prostaglandins 15d-PGJ2, PGE2 and PGD2 were purchased from Cayman Chemicals, while epoxycyclopentenone (EC), cyclo-epoxycyclopentenone (cEC), EC-reduced were synthetized as previously described [[31] (link), [32] (link), [33] (link)]. The antioxidant N-acetylcysteine (NAC) and the pan-caspase inhibitor Q-VD-OPh (QVD) were purchased from Sigma-Aldrich, whereas the necroptosis inhibitor Nec-1s was from Merck Millipore.
4
Nrf2 Modulation Using Brusatol and tBHQ
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Brusatol (BRU), a Nrf2 inhibitor, was obtained from Tauto (Shanghai, China). Dimethyl sulfoxide (DMSO) and tert-butylhydroquinone (tBHQ), a Nrf2 inducer, were obtained from Sigma–Aldrich (St. Louis, MO, USA).
5
Synthesis of Novel Naphthalene Analogues
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SFN, Carbonyl cyanide-p-trifluoromethoxyphenylhydrazone (FCCP), dimethylfumarate (DMF), curcumin and tert-Butylhydroquinone (TBHQ) were purchased from Sigma-Aldrich. PMI (compound 1 ) and analogues (compounds 2 –4 ) were prepared as described previously13 (link). Compounds 5 and 6 were synthesized starting from 1-nitronaphthalene using adaptations of the published procedures18 (link), 19 (link).
6
Hepatoprotective Evaluation of Tert-butylhydroquinone
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Tert-butylhydroquinone (purity: >97.0%) was purchased from Sigma Aldrich (St. Louis, MO, USA). CCl4 (purity>98%) was purchased from Acmec biochemical (Shanghai, China). The kits for measurement levels of alanine transaminase (ALT), reduced glutathione/oxidized glutathione (GSH/GSSG) level, aspartate transaminase (AST), malondialdehyde (MDA), and olive oil were purchased from the Nanjing Jiancheng Institute of Biotechnology (Nanjing, China). enzyme-linked immunosorbent assay (ELISA) kits for detection of mice interleukin-6(IL-6), interleukin-1β (IL-1β), interferon-γ (IFN-γ), monocyte chemotactic protein 1 (MCP-1) and Tumor necrosis factor-α (TNF-α) were purchased form Dakewe Bioengineering Co., Ltd. Fetal bovine serum and RPMI-1640 medium were obtained from Gibco Life Technologies (Carlsbad, CA, USA). Annexin V-FITC Apoptosis Detection Kit, Caspase-3 level detection kit and 2,7-Dichlorofluorescein diacetate (DCFH-DA) was purchased from Beyotime Biotechnology (Shanghai, China). Anti-heme oxygenase-1 (HO-1), anti-Nrf-2 and b-actin were obtained from Cell Signaling Technology (Danvers, Mass). TUNEL Apoptosis Assay Kit (ROCHE In situ Cell Death Detection Kit, TMR red) was purchased from Roche. All other chemicals were of the highest commercial grade.
7
Chemical Stress Induction Assay
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Tunicamycin (TM) and thapsigargin (TG) were purchased from EMD Millipore Corporation (Billerica, MA, United States); Hydroquinone (HQ), tert-ButylHydroquinone (tBHQ), and cycloheximide (CHX) were purchased from Sigma-Aldrich (St. Louis, MO, United States); MG132 was purchased from Cayman Chemical (Ann Arbor, MI, United States). 4μ8C was provided by Dr. David Ron (University of Cambridge Metabolic Research Laboratories and National Institute for Health Research Cambridge Biomedical Research Centre, United Kingdom); Quinotrierixin (QT) was a kind gift from Dr. Etsu Tashiro (Keio University, Japan).
8
HPLC Analysis of Vitamins A and E
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High-performance liquid chromatography (HPLC) grade solvents, methanol, acetonitrile, n-hexane, acetone, petroleum ether, dichloromethane, cyclohexane, isopropanol, diethyl ether, and ethyl acetate were purchased from Fluka and Riedel (Munich, Germany) and were used in all procedures without further purification. Tert-butylhydroquinone was used as an antioxidant and was obtained from Sigma–Aldrich (Munich, Germany). K2HPO4 (extra pure), and trifluoroacetic acid were purchased from Merck (Darmstadt, Germany). Karl Fischer titration reagents were from Scharlau (Sentmenat, Spain) High purity water was obtained through a Milli-Q water purification system (Millipore, Bedford, MA, USA) and was used in all procedures. Vitamins A and E (all–trans-retinol, retinyl palmitate, α-tocopherol and γ-tocopherol) were of analytical reagent grade. All the vitamins were supplied by Sigma–Aldrich.
9
Dissecting Insulin Signaling Pathways
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Tert-Butylhydroquinone (TBHQ), streptozocin (STZ), insulin and HClO were provided by (Sigma, St. Louis, MO, USA).
Fetal bovine serum (FBS) and Dulbecco's Modified Eagle's Medium (DMEM) were provided by Gibco BRL (Gibico, Grand Island, NY, USA). The bicinchoninic acid (BCA) protein assay, hematoxylin and eosin (HE) and Hoechst staining kits were provided by Beyotime (Beijing, China). Antibodies targeting p-AKT, AMPKα2, glycogen synthase kinase-3 β (GSK3 β), and glucose transport-4 (GLUT4) were provided by Abcam (Cambridge, MA, USA). Anti-phosphatidylinositol 3-kinase (PI3K) and anti-AKT antibodies were provided by Sangon (Shanghai, China). Antibodies directed against p-PI3K were provided by Cell Signaling Technology (Danvers, MA, USA). Antibodies targeting β-actin were provided by Beyotime (Beijing, China). PRKAA2 siRNA and riboFECT™ CP transfection kit were provided by Guangzhou RiboBio (Guangzhou, China).
Fetal bovine serum (FBS) and Dulbecco's Modified Eagle's Medium (DMEM) were provided by Gibco BRL (Gibico, Grand Island, NY, USA). The bicinchoninic acid (BCA) protein assay, hematoxylin and eosin (HE) and Hoechst staining kits were provided by Beyotime (Beijing, China). Antibodies targeting p-AKT, AMPKα2, glycogen synthase kinase-3 β (GSK3 β), and glucose transport-4 (GLUT4) were provided by Abcam (Cambridge, MA, USA). Anti-phosphatidylinositol 3-kinase (PI3K) and anti-AKT antibodies were provided by Sangon (Shanghai, China). Antibodies directed against p-PI3K were provided by Cell Signaling Technology (Danvers, MA, USA). Antibodies targeting β-actin were provided by Beyotime (Beijing, China). PRKAA2 siRNA and riboFECT™ CP transfection kit were provided by Guangzhou RiboBio (Guangzhou, China).
10
Quantification of Drug Transporter Expression
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Rhodamine 123, probenecid, verapamil, fumitremorgin C, rifampicin, phenobarbital, chenodeoxycholic acid (CDCA) and tert-butylhydroquinone (tBHQ) were provided by Sigma-Aldrich (Saint-Quentin Fallavier, France), whereas carboxy-2,7-dichlorofluorescein (CF) diacetate and Hoechst 33342 were from Life Technologies (Saint Aubin, France) and 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) from Cambridge Isotope Laboratories (Cambridge, UK). [3H(G)] taurocholic acid (sp. act. 5.0 Ci/mmol), [6,7-3H(N)] estrone-3-sulfate (E3S) (sp. act. 54.0 Ci/mmol) and [1-14C] tetra-ethylammonium (TEA) (sp. act. 3.5 mCi/mmol) were supplied by Perkin-Elmer (Boston, MA, USA). Mouse monoclonal antibodies against P-gp (clone C219), MRP2 (clone M2III-6) and MRP3 (ABCC3) (clone M3II-9) were from Alexis Biochemicals (Lausen, Switzerland), whereas rabbit antibody against the p38 mitogen-activated protein kinase (MAPK) was provided by Santa Cruz Biotechnology (Dallas, TX, USA). Mouse monoclonal antibody against MRP4 (ABCC4) and rabbit polyclonal antibody against MRP5 (ABCC5) were purchased from Abcam (Paris, France). All other chemicals and reagents were commercial products of the highest purity available.
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