Prl tk renilla luciferase internal control vector

Induction of NFκB transcriptional activity was evaluated using a luciferase reporter gene driven by κ enhancer, pNFκB (Clontech, Palo Alto, CA). Raw 264.7 cells were transfected with the luciferase reporter construct together with pRL-TK Renilla luciferase internal control vector (Promega, Madison, WI) using Lipofectamine LTX and Plus reagent (Invitrogen) according to the manufacturer's instructions. After 24 h, cells were incubated with 10 μg protein/ml OxLDL in the presence or absence of 75 μg/ml C1q for a further 24 h. Cells were then harvested and the firefly luciferase activity and the Renilla luciferase activity were measured by the Dual Luciferase reporter assay system using a TD 20/20 luminometer (Promega). The firefly luciferase activity was normalized to the Renilla luciferase activity to control for variability in transfection efficiency and to calculate the relative luciferase activity units (RLU).