3 4 5 dimethylthiazol 2 yl 5 3 carboxymethoxyphenyl 2 4 sulfophenyl 2h tetrazolium inner salt mts assay kit

Aminoguanidine hydrochloride (AG), MGO (40% aqueous solution), bovine serum albumin (essentially fatty acid-free), acetic anhydride, pyridine, magnesium sulfate (MgSO₄), d-gluconolactone, N-acetyl-glycyl-lysine methyl ester acetate salt (G.K. peptide), d-ribose, d-glucose, sodium azide (NaN₃), sodium bicarbonate, methanol (MeOH), anhydrous acetone, prenyl bromide, potassium carbonate (K₂CO₃), Griess reagent system, and CS were purchased from Sigma-Aldrich (St. Louis, MO, USA). The 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt (MTS) assay kit was purchased from Promega (Promega Co., Madison, WI, USA). All solvents and other reagents used in this study, unless otherwise specified, were analytical grade and purchased from Sigma-Aldrich (St. Louis, MO, USA).

7,8-DHF was purchased from Sigma-Aldrich (St. Louis, MO, USA) and dissolved in dimethyl sulfoxide (DMSO, Sigma-Aldrich). Dulbecco’s modified Eagle medium (DMEM), fetal bovine serum (FBS), bovine calf serum (BCS), penicillin-streptomycin, and 2-NBDG were purchased from Invitrogen (Carlsbad, CA, USA). The 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxy-methoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, Inner Salt (MTS) assay kit was obtained from Promega (Madison, WI, USA). The enzyme-linked immunosorbent assay (ELISA) kits for TNF-α, IL-6, MCP-1, and adiponectin were obtained from R&D Systems (Minneapolis, MN, USA). Nuclear and cytoplasmic extraction reagents and the assay kit for NEFA were provided by Thermo Fisher Scientific (Rockford, IL, USA) and FUJIFILM Wako Pure Chemical Corporation (Chuo-Ku, Osaka, Japan), respectively. Antibodies specific for phospho-c-Jun N-terminal kinases (p-JNK), total-JNK (t-JNK), phospho-Akt (Ser473), Akt, and NF-κB were purchased from Cell Signaling Technology (Danvers, MA, USA). The antibody for lamin B was purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA), and antibodies for α-tubulin and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) were from Ab Frontier (Seoul, Korea). All other chemicals were obtained from Sigma-Aldrich.