Luciferase Reporter Assay for IFN Signaling

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.