Nrf2 transcriptional activity

Nrf2‐dependent antioxidant response element (ARE) luciferase reporter assay was conducted as described previously (Gan et al, 2013). Briefly, cells were seeded in a 24‐well culture plate and transfected with 0.15 μg of pGL3‐ARE firefly luciferase reporter plasmid and 0.1 µg pRL‐CMV. 24 h post‐transfection, cells were lysed, and the luciferase activities were determined using the Dual‐Luciferase® Reporter Assay System (Promega). Results were expressed as firefly luciferase activity normalized to renilla luciferase activity.