Fetal bovine serum fbs s11150

Human astroglial SVG-A derived cells (a kind gift from Walter J. Atwood, Brown University, Providence, RI) were grown at 37 °C and 5% CO₂ in Minimum Essential Medium (MEM) (10-010-CV; Corning) supplemented with 10% heat-inactivated fetal bovine serum (FBS, S11150; Atlanta Biologicals), penicillin, and streptomycin (1406-05-9; VWR International). African Green Monkey kidney epithelial MA104 cells (a kind gift from Siyuan Ding, Washington University in St. Louis, St. Louis, MO) were grown at 37 °C and 5% CO₂ in Medium 199 supplemented with 10% heat-inactivated FBS. Vero C1008 (Vero 76, clone E6, Vero E6) [American Type Culture Collection (ATCC) CRL-1586] cells were cultured in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% FBS, and penicillin and streptomycin. Vero CCL-81 (ATCC CCL-81) cells were maintained in DMEM supplemented with 10% FBS, 10 mM Hepes pH 7.4, 1% Glutamax, and penicillin/streptomycin.

Huh7 (hepatocyte-derived carcinoma cell line) was purchased from the American Type Culture Collection (ATCC; Manassas, VA, USA). Dulbecco’s modified eagle medium (DMEM; 11965-092) was manufactured by Gibco (Waltham, MA, USA). Fetal bovine serum (FBS; S11150) was produced by Atlanta Biologicals (Flowery Branch, GA, USA). Penicillin-streptomycin (P0781) and sodium pyruvate (S8636) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Huh7 cells were cultured in DMEM which supplemented with 10% FBS (v/v), 1 mM sodium pyruvate, 100 units/mL penicillin, and 100 µg/mL streptomycin at 37 °C containing 5% CO₂ in the humidified atmosphere. Transfections with each plasmid were performed using FuGENE 6 per the manufacturer’s protocols. For each well, 1 × 10⁵ Huh7 cells were distributed into a 100 mm culture dish and incubated at 37 °C for 24 h. The transfection mixture was prepared with containing 5 µg of each cDNA, 25 µL of FuGENE 6 and 10 mL Opti-MEM reduced serum medium (31985-070, Gibco) at room temperature for 15 min. When the cells turned into adhesive, the growth medium was replaced with the transfection mixture and cells were incubated for additional 24 h at 37 °C.