Eagle s minimal essential medium emem

2-Methoxyestradiol (2ME) was purchased from Fraken Biochem Co., Ltd., (Qingdao, China) with purity more than 98%; ethanol, chloroform, and dimethyl sulfoxide (DMSO) were obtained from Sigma-Aldrich (St. Louis, MO, USA). Tween 80, cumin oil, and propylene glycol were purchased from Sigma-Aldrich (St. Louis, MO, USA). Thiazolyl Blue Tetrazolium Bromide (MTT) was obtained from Sigma-Aldrich Inc. (St. Louis, MO, USA). Eagle’s Minimal Essential Medium (EMEM) was obtained from Sigma-Aldrich Co. (St. Louis, MO, USA); TrypLE™ Express Enzyme (1X), phosphate-buffered saline (PBS), penicillin/streptomycin, and fetal bovine serum (FBS, Qualified) were purchased from Gibco (Grand Island, NY, USA). Doxorubicin (DOX) was purchased from MCE (Med Chem Express, Monmouth Junction, South Brunswick, NJ, USA).

5-Aminolevulinic acid hydrochloride (Lot number: HCL-KK08-04-1-1, purity; 99.8%) was obtained from Cosmo Oil Co., Ltd, (Tokyo, Japan). SFC was purchased from Komatsuya Corporation (Osaka, Japan). DL-Dithiothreitol, Oil Red O, sodium orthovanadate (Na₃VO₄), Tween 20, Eagle’s minimal essential medium (EMEM), and 100 × antibiotic antimycotic solution (ABAM) were purchased from Sigma-Aldrich Japan (Tokyo, Japan). Sodium fluoride (NaF), sodium deoxycholate, l-ascorbic acid phosphate magnesium salt n-hydrate, isopropanol, and Dulbecco’s modified Eagle’s medium (DMEM) with 4500 mg/L glucose (DMEM-high glucose) and 1000 mg/L glucose (DMEM-low glucose) were purchased from Wako Pure Chemicals (Tokyo, Japan). Fetal bovine serum (FBS) and Calf bovine serum (CBS) were purchased from BioWest (Nuaillé, France) and the American Type Culture Collection (ATCC, Manassas, VA, USA), respectively. Insulin (10 μg/mL human recombinant) was purchased from Life Technologies Japan (Tokyo, Japan). Halt Protease Inhibitor Cocktail was purchased from Thermo-Scientific (Waltham, MA, USA). High-fat diet (HFD) (60 kcal% fat, D12492) was purchased from Research Diet Inc. (New Brunswick, NJ, USA).

Analytical grade (to enable its use without further purification) rhodamine 123 (R123); sodium dodecyl sulfate (SDS); 3-(4.5-dimethylthiazol-2-yl)-2.5-diphenyltetrazolium bromide (MTT); Dulbecco’s Modified Eagle’s medium–high glucose (DMEM) with 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin mixture; resazurin sodium salt; trypsin-EDTA solution; allantoin; and tariquidar and dimethyl sulfoxide (DMSO) were acquired at Sigma-Aldrich (St. Louis, MO, USA). Doxorubicin hydrochloride was acquired from Teva Pharmaceuticals (Petah Tikva, Israel). Eagle’s Minimal Essential Medium (EMEM, Sigma-Aldrich) containing 4500 mg/L glucose, supplemented with a non-essential amino acid (NEAA) mixture (Sigma-Aldrich); a selection of vitamins and 10% heat-inactivated FBS; 2 mM L-glutamine (Sigma-Aldrich); 1 mM Na-pyruvate (Sigma-Aldrich); nystatin (Sigma-Aldrich); a penicillin-streptomycin mixture at concentrations of 100 U/L and 10 mg/L; RPMI 1640 medium (Sigma-Aldrich), supplemented with 10% FBS; 2 mM L-glutamine; 1 mM Na-pyruvate; 100 mM HEPES (Sigma-Aldrich); nystatin; and a penicillin-streptomycin mixture at concentrations of 100 U/L and 10 mg/L were used in the biological evaluation. Pgp-Glo™ Assay Systems (Promega), and an Annexin V-FITC Apoptosis Detection Kit were used (Calbiochem, EMD Biosciences. Inc. La Jolla, CA, USA).