Nicotine adenine dinucleotide phosphate reduced salt

HPLC-grade Acetonitrile and water were purchased by Panreac (Madrid, Spain). Standards of kaempferol, myricetin, quercetin, rutin, caffeic, chlorogenic, and protocautechic acids of HPLC-grade were purchased by Sigma-Aldrich (Madrid, Spain). We also acquired CCCP from Sigma (C2759).
Tert-butylhydroperoxide (t-BOOH), glutathione reductase (GR), reduced (GSH) and oxidized (GSSG) glutathione, dichlorofluorescin (DCFH), o-phthaldialdehyde (OPT), nicotine adenine dinucleotide phosphate reduced salt (NADPH), 2,4-dinitrophenylhydrazine (DNPH), H₂O₂, 1,1,3,3-tetraethoxypropane (TEP), gentamicin, penicillin G, and streptomycin were purchased from Sigma Chemical Co. (Madrid, Spain), and rapamycin was purchased from Merck (number 553,210). Acetonitrile, methanol of HPLC grade, dimethyl sulfoxide (DMSO) of analytical grade, sodium hydroxide, sodium chloride, di-sodium hydrogen phosphate anhydrous, potassium di-hydrogen phosphate, as well as formic, hydrochloric, perchloric, and sulfuric acids were acquired from Panreac (Barcelona, Spain). Bradford reagent was from BioRad Laboratories S.A. DMEM culture media and fetal bovine serum (FBS) were from Cultek (Madrid, Spain). All other reagents were of analytical quality.

Dulbecco’s modified eagle’s medium (DMEM) and fetal bovine serum (FBS) were from Bio Whitaker Europe (Lonza, Madrid, Spain). DHCA, DHFA, HHA, gentamicin, penicillin, streptomycin, o-phthaldialdehyde (OPT), GR, GSH and oxidised (GSSG) glutathione, nicotine adenine dinucleotide (reduced) (NADH), nicotine adenine dinucleotide phosphate reduced salt (NADPH), 20,70-dichlorofluorescein diacetate (DCFHDA), 2,4-dinitrophenylhydrazone (DNPH), β-mercaptoethanol, guanidine, dithiothreitol (DTT), ethylenediaminetetraacetic acid (EDTA), dimethylsulphoxide (DMSO), t-BOOH, and sodium dodecyl sulphate (SDS) were purchased from Sigma-Aldrich (Madrid, Spain). Bradford reagent was acquired from Bio-Rad (Madrid, Spain). Recombinant murine TNF-α was from PreproTech (Tebu-bio, Madrid, Spain). IL-6 ELISA kit was from R&D Systems (Materlab, Madrid, Spain). The Pro Human Cytokine, Chemokine, and Growth Factor Assay (group I) Multiplex kit was purchased from Bio-Rad (Madrid, Spain). Cell culture dishes were from Falcon (Cajal, Madrid, Spain). Human hepatoma HepG2 cells were a kind gift from Dr. Paloma Martin-Sanz (Instituto de Investigaciones Biomédicas Alberto Sols, CSIC, Madrid, Spain). All other chemicals were of analytical grade.

Nicotine adenine dinucleotide reduced salt, nicotine adenine dinucleotide phosphate reduced salt (NADPH), 2′,7′-dichlorofluorescin diacetate (DCFH), reduced glutathione (GSH), glutathione reductase (GR), tert-butylhydroperoxide, o-phthaldehyde, streptavidin–biotin conjugated horseradish peroxidase (HRP), 3,3′-diaminobenzidine (DAB), and glucose were purchased from Sigma Chemical (Madrid, Spain). Antiproliferative cell nuclear antigen (PCNA; PC-10) was purchased from Lab Vision Corporation and Bionova- Científica SL. Anti-SIRT1 (sc-74465), anti-occludin (sc-5562), anti-ZO-1 (sc-33725), anti-TNF-α (sc-52746), and anti-IL6 (sc-57315) were purchased from Santa Cruz Biotechnology (Quimigen, Madrid, Spain). Antiphospho-Nrf2 (#12811) was purchased from Signalway antibody (Quimigen, Madrid, Spain). Terminal transferase recombinant, biotin-16-dUTP, and proteinase K were purchased from Roche Applied Science (Roche Diagnostic, GMbH, Madrid, Spain).