HEK 293T cells were transfected with the indicated TNFR1 construct along with a Renilla luciferase construct (pGL4.74, Promega) and the luc2/NFκB/RE reporter (pGL4.32, Promega). Cells were analyzed for firefly normalized to Renilla luciferase activity using the Dual Luciferase Reporter Assay System (Promega). IL-8 levels in culture supernatant were measured by immunoassay using the Human CXCL8/IL-8 Quantikine® ELISA kit (R & D Systems), according the manufacturer’s instructions.
