EMSA and supershift analysis of the rat GCLC AP-1 and NF-κB sites. WT cells were treated with TBH (60 μM for 8 h) or vehicle control and subjected to EMSA with supershift analysis for the AP-1 site at −356 or the NF-κB site at −378. Supershift analysis was performed using antibodies directed against c-Jun, Nrf1, and Nrf2 for the AP-1 site (A) and p50, Nrf1, and Nrf2 for the NF-κB site (B). Note that supershift occurred only with anti-c-Jun antibodies for the AP-1 site and anti-p50 antibodies for the NF-κB site. As a positive control, TBH treatment induced Nrf1 and Nrf2 binding to the ARE site of the mouse GCLM (C). Arrows in panel C point to the Nrf1 and Nrf2 supershifts.