Gentamicin

Human coronary artery smooth muscle cells (CASMC, #CC-2583; Clonetics, Walkersville, MD) were obtained at passage 3 and used in passages not higher than 10. CASMC were grown in smooth muscle cell basal medium (SmBM, #CC-3182; Clonetics, Walkersville, MD), including 0.5 µg/ml hEGF, 5 mg/ml insulin, 1 µg/ml bFGF (basic fibroblast growth factor), 50 mg/ml gentamicin and 5% FBS; (#CC-4149; Clonetics, Walkersville, MD) at 37°C in 5% CO₂. A differentiated CASMC phenotype was induced by serum depletion [4] (link) by smooth muscle cell basal medium (SmBM, #CC-3182; Clonetics, Walkersville, MD), including 0.5 µg/ml hEGF (human epidermal growth factor), 5 mg/ml insulin, 1 µg/ml bFGF and 50 mg/ml gentamicin w/o FBS (fetal bovine serum) at 37°C in 5% CO₂ for 12 h, 24 h, 36 h, 48 h, 60 h and 72 h.

Dulbecco’s Modified Eagle Medium (DMEM), heat-inactivated low-endotoxin fetal bovine serum (FBS), and gentamicin purchased from Cambrex (Walkersville MD, USA) for tissue culture. All other cancer cell lines were purchased from American Type Culture Collection (Manassas, VA, USA). Most of these following compounds (thiostrepton, ampicillin, dexamethasone, 2-methoxyestradiol, (−) riboflavin, ascorbic acid, amiloride-HCL, (−) Corey lactone, and quinine sulphate) were purchased from Sigma-Aldrich Chemical Co. (St. Louis, MO, USA), and quercetin was purchased from Alfa Aesar (Johnson Matthey Co. Lancastor, UK). The 50% purified δ-tocotrienol fraction from annatto seeds was purchased from American River (Boston, MA, USA). RNeasy mini kit was purchased from QIAGEN Sciences (Germantown, MD, USA). DeltaGold (125 mg soft gels) from annatto seeds (composition 90% δ-tocotrienol + 10% γ-tocotrienol) was obtained as a gift by American River Nutrition, Inc. (Hadley, MA. USA). The hepatitis C antibody test kit was purchased from Sigma Chemical Co., St. Louis, USA. The second kit for diagnosing hepatitis C test is RNA (PCR) test. Pure total mRNA was obtained from the EDTA treated fresh whole blood using total RNA purification kit # 17200 (NORGEN Bioteck Corporation, Thorold, ON, Canada).