BD Vacutainer EDTA (purple capped) blood collection tubes were purchased from Becton Dickinson and Company. DNeasy blood & tissue kit and RNase (100 mg mL–1) were acquired from Qiagen Ltd. TaqMan Drug Metabolism SNP Genotyping Assays and TaqMan Universal Master Mix II without uracil‐N‐glycosylase (UNG) were purchased from Thermo Fischer Scientific. Sulforaphane (CAS 4478‐93‐7) (purity > 98%) was purchased from LKT Laboratories. The internal standard N‐butylthiocarbamoyl cysteine (B‐ITC) and Sulforaphane conjugates including Sulforaphane‐glutathione, Sulforaphane‐cysteine‐glycine, Sulforaphane‐cysteine, Sulforaphane‐N‐acetylcysteine (‐NAC), and erucin‐NAC were synthesized as published in.11 The internal standard sinigrin for glucoraphanin was purchased from Sigma–Aldrich. Blank plasma from healthy participants with the same diet restriction described in ‘Study design’ was ordered from Sera Laboratories International Ltd to make standard curves. DEAE Sephadex A25 and SP Sephadex C25 were obtained from Amersham Biosciences. Glucoraphanin (CAS 21414‐41‐5) (purity ≥ 95%) and glucoerucin (CAS 21973‐56‐8) (purity ≥ 97%) were purchased from Cayman Chemical and from Carl Roth, respectively. All other chemicals were purchased from Sigma–Aldrich.
Sulforaphane
Manufactured by Merck Group
Sourced in United States, France, China
Sulforaphane is a chemical compound found naturally in certain vegetables, such as broccoli and Brussels sprouts. It is a product developed by Merck Group for use in laboratory research settings.
Automatically generated - may contain errors
Lab products found in correlation
Dimethyl sulfoxide (dmso), by Merck Group (10 mentions)
Quercetin, by Merck Group (9 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (9 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (8 mentions)
Mg132, by Merck Group (6 mentions)
Curcumin, by Merck Group (5 mentions)
N acetyl l cysteine, by Merck Group (4 mentions)
Cycloheximide (chx), by Merck Group (4 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (4 mentions)
Dimethyl fumarate, by Merck Group (3 mentions)
Allyl isothiocyanate, by Merck Group (3 mentions)
Caffeic acid, by Merck Group (3 mentions)
Brusatol, by Biosynth (3 mentions)
Luciferase assay system, by Promega (3 mentions)
Hemin, by Merck Group (3 mentions)
Pen strep solution 100x, by Thermo Fisher Scientific (3 mentions)
Resveratrol, by Merck Group (3 mentions)
Nicotinamide, by Merck Group (3 mentions)
Protease inhibitor cocktail, by Merck Group (3 mentions)
Actinomycin d, by Merck Group (3 mentions)
98 protocols using sulforaphane
1
Quantification of Sulforaphane and Metabolites
2
Pharmacological Agents for In Vitro Studies
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2-(4-Morpholinethyl)-1-phenylcyclohexanecarboxylate hydrochloride (PRE-084), 1-(3,4-Dimethoxyphenethyl)-4-(3-phenylpropyl)piperazine) dihydrochloride (SA4503), 1-isothiocyanato-4-methylsulfinylbutane (sulforaphane) and tunicamycin were from Sigma-Aldrich (Saint-Quentin-Fallavier, France). 4-Methoxy-3-N,N-dipropylbenzeneethanamine (NE-100) was purchased from Tocris (Tebu-Bio, Le-Perray-En-Yvelines, France). PRE-084, SA4503 and NE-100 were solubilized in water and tunicamycin (10 mg/ml) and sulforaphane (4 mg/ml) in pure DMSO (Sigma-Aldrich).
3
Modulation of THP-1 Macrophages by Stimuli
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THP-1 cells (ATCC) were cultured in RPMI 1640 (Thermo) supplemented with 10% FCS (R&D) and 1% Pen/Strep (Thermo). LUCAT1−/− THP-1 have been generated previously (10 (link)). Cells were differentiated into macrophage-like cells using 10 ng/mL phorbol-12-myristate acetate (PMA, Sigma) for 24 h, followed by media change and resting in PMA-free medium for another 24 h before stimulation. Cells were treated with 200 ng/mL LPS from E. coli 0111:B4 (Invivogen), Sendai virus Cantrell strain (Charles River Laboratories), 10 ng/mL IFN-α 2b (Gemini), 100 µM to 250 µM 4-octyl itaconate, 250 µM di-methyl itaconate, 5 µM sulforaphane (all Sigma), or 500 nM diABZI-4 (GSK) (19 (link)).
4
Bardoxolone Methyl and Sulforaphane Assays
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Bardoxolone methyl was purchased from Sigma-Aldrich (SMB00376) or MedChemExpress (HY-13324). Stock concentrations of 100 and 400 µM were prepared in dimethyl sulfoxide (DMSO), and then diluted 1:1000 into the cell culture medium. Sulforaphane was purchased from Sigma-Aldrich (S4441). Stock concentrations of 2 and 5 mM were prepared in DMSO, and then diluted 1:1000 into the cell culture medium. For the assays combining treatment with HSV-1 infection, half of the medium was removed, virus inoculum at an MOI of 1 was added to the remaining medium, and half an hour later the cells were washed with warm PBS, the conditioned medium added back to the cells with Bardoxolone methyl or Sulforaphane supplied at the indicated concentrations.
5
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.
6
Phytochemical Screening and Antioxidant Evaluation
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The following chemicals were used: methanol, acetonitrile, isopropanol (HPLC grade, Merck, Darmstadt, Germany), formic acid (>96.0%), 2,2-azinobis-(ethyl-2,3-dihydrobenzothiazoline-6-sulphonic acid) diammonium salt-ABTS, 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid-Trolox, caffeic acid, ferulic acid, sinapic acid, quercetin, kaempferol, indole-3-carbinol, indole-3-acetonitrile, 3,3′-diindolylmethane, N-acetyl-l -cysteine, allyl isothiocyanate, sulforaphane (Sigma Aldrich, St. Louis, MO, USA), and glucotropaeolin (AppliChem, Darmstadt, Germany).
7
Dimethyl Fumarate and Cell Signaling
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Dimethyl fumarate was obtained from Sigma-Aldrich company (#50744, Sigma-Aldrich, St. Louis, USA). Tween-20 (#P1379), LPS from Escherichia coli 0111:B4 (#L2690), Nrf-2 agonist Sulforaphane (#S6317) and ROS inhibitor NAC (#1009005) were also from Sigma.
8
Antioxidant and Anti-inflammatory Assays
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Folin-Ciocalteu phenol reagent, gallic acid, dehydroascorbic acid, 2,2′-azobis-2-methyl-propanimidamide, dihydrochloride (AAPH), fluorescein sodium salt (FL), 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (Trolox), and sulforaphane were purchased from Sigma-Aldrich Co. (St. Louis, MO, USA). Dulbecco’s modified Eagle’s medium (DMEM), fetal bovine serum (FBS), and penicillin/streptomycin were purchased from Gibco BRL (Life Technologies Inc., Grand Island, NY, USA). LPS (Escherichia coli O111:B4) was obtained from Sigma-Aldrich Co. iNOS was purchased from Calbiochem (San Diego, CA, USA). IκB was purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). The acetonitrile was HPLC grade, and all other chemicals were analytical grade.
9
Luciferase Assay for Nrf2 Activation
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Luciferase reporter assay was conducted on HepG2-ARE cells as described [18 (link), 20 (link)]. The cells were treated with samples for 12 h after serum starvation (0.5% FBS, 12 h). The luciferase activity, which corresponded to the ARE activity, was measured using a luciferase assay system (Promega) according to the manufacturer's instruction. Sulforaphane (Sigma-Aldrich), an isothiocyanate, was used as an ARE activator. Brusatol (Carbosynth Ltd., Newbury, Berkshire, UK), a quassinoid, was used as a specific inhibitor of the Nrf2 pathway [21 (link)]. The luminescence of the assay was detected and calibrated on total protein amounts. The data were then normalized against the control values.
10
Acrylamide and Sulforaphane Administration
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Acrylamide (lot #A9099, purity > 99%) and sulforaphane (lot #3200372) were purchased from Sigma Aldrich (St. Louis, MO, USA) and LKT Laboratories Inc. (St. Paul, MN), respectively. Acrylamide was freshly prepared at the beginning of each week by dissolving in a G-10 ion exchange cartridge (Organo Co., Tokyo, Japan) filtered drinking water, stored at 4 degrees Celsius and administered every day in autoclaved bottles [43 (link)]. sulforaphane was prepared by dissolving the stock solution in normal saline just before treatment.
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