Prl cmv encoding renilla luciferase

Cells were transfected using Lipofectamine 2000 (Invitrogen) with pRL-CMV encoding Renilla luciferase (Promega) and luciferase reporter constructs for either gamma-interferon activation site (GAS) (Panomics, Fremont, CA, USA) or IFN-stimulated regulatory element (ISRE) (kindly provided by Dr Izortze Santin, University of the Basque Country, Spain). After recovery, cells were treated with either IFNα for 2 h or IFNγ for 24 h (30 (link)). Luciferase activity was measured in a POLASTAR plate reader (BMG Labtech) using the Dual-Luciferase Reporter Assay System (Promega) and corrected for the luciferase activity of the internal control plasmid, i.e., pRL-CMV.

Branched polyethylenimine (PEI, Mw ~ 25 kDa), ethylenediamine (EDA, 99.5%), methyl acrylate (MA, 99.0%), 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDAC, 98%), 1,1′-carbonyldiimidazole (CDI, 97%), 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyl tetrazolium bromide (MTT), fluorescein diacetate (FDA, >98%), propidium iodinate (PI, >98%), D-mannitol (>99%), and 5-fluorocytosine (5-FC) were obtained from Sigma-Aldrich Chemical (St. Louis, MO, USA). Female Balb/c-nu mice were purchased from Institute of Laboratory Animal Sciences (ILAS). The plasmid DNAs (pDNA), including pRL-CMV encoding renilla luciferase (Promega Co., Cergy Pontoise, France), pEGFP-N1 encoding enhancedgreen fluorescent protein (EGFP) (BD Biosciences, San Jose, CA, USA), and pAdTrack-CMV-CD (pCMV-CD) encoding Escherichia coli cytosine deaminase (ECD) were amplified in Escherichia coli and purified according to the supplier’s protocol (Qiagen GmbH, Hilden, Germany). The quality and concentration of the purified pDNA were assessed by measuring its absorption at 260 nm and 280 nm, and by agarose gel electrophoresis. HepG2 and COS7 cell lines were purchased from the American Type Culture Collection (ATCC, Rockville, MD, USA).