Mt35011cv

PCCL3, a normal TSH-dependent rat thyroid follicular cell line, was grown in Coon’s Media: Nutrient Mixture F-12 Ham (Coon’s modification; Sigma-Aldrich, F6636) supplemented with 2.68 g/L sodium bicarbonate, 5% fetal bovine serum (FBS; Corning, MT35011CV), 1% penicillin/streptomycin, 2 mM L-glutamine, insulin (1 μg/mL), apo-transferrin (5 μg/mL), hydrocortisone (1 nM), and thyroid-stimulating hormone (TSH 1 mIU/mL). The rat hepatoma clonal cell line HC1 (kindly provided by Dr. Elliot Ross, University of Texas Southwestern Medical Center) was grown in DMEM (Corning, 10013CV) supplemented with 10% FBS, penicillin (100 IU/L), and streptomycin (100 mg/L), and L-glutamine. Cells were grown to ~90% confluency before passaging every 2 to 4 d at 37 °C in 5% CO2, 95% humidified air. Transient transfections were performed using Lipofectamine 3000 Transfection kit (Invitrogen) for 24 h. NLS-bPAC-nLuc stable cell lines were generated by lentiviral infection with a multiplicity of infection (MOI) of 80 and 5 μg/mL of polybrene and selected with puromycin as described before (52 (link)).

The standard nonagitation aggregation assay was used to assess activity of 0 to 30% (v/v) of FBS (#MT35011CV, Corning) or lipoprotein-deficient FBS (#S5394, Sigma-Aldrich).