2.10. HIF-1α Luciferase Assay

RAW 264.7 cells were transfected with luciferase reporter plasmid combined with a HIF-1α-Luc reporter for 48 h, treated with nutlin-3a for 2 h, stimulated with LPS for 48 h, and then lysed, followed by analysis of the reporter activity using a luminometer 1420 luminescence counter (Perkin Elmer, Waltham, MA, USA). The relative luciferase activity was determined by the ratio of firefly luciferase activity to renilla luciferase activity.