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. Author manuscript; available in PMC: 2017 Dec 1.
Published in final edited form as: Free Radic Biol Med. 2016 Oct 4;101:116–128. doi: 10.1016/j.freeradbiomed.2016.10.003

Figure 4.

Figure 4

Mechanisms of Nrf2 induction by WA. A) Nqo1 transcript induction in Keap1-disrupted (K0), WT, Nrf2-disrupted (N0) and Keap1 & Nrf2 double-disrupted (K0N0) MEF with graded doses of WA (0–3 μM), 20 hours post-treatment. B) Relative luciferase activity in Keap1 & Nrf2 double-disrupted MEF transfected with Nqo1-ARE luciferase reporter vector and either 5 ng pCMV Nrf2 alone or with 5 ng pCMV Nrf2 and 2.5 ng pCMV WT Keap1/pCMV C151S Keap1. The reconstituted cells were then treated with DMSO (Veh), 3 μM WA or 10 μM sulforaphane (SFN) for 20 hours. Transfection efficiencies were quantified by normalization to Renilla luciferase plasmid vector. Statistical comparisons: [*compared to mock control. **compared to + Nrf2 + Veh control. #compared to + Nrf2 + WT Keap1 + Veh control. ##compared to + Nrf2 + WT Keap1 + SFN. $compared to + Nrf2 + C151S Keap1 + Veh control]. Effects of LY294002 pre-treatment on WA-mediated Nqo1 induction in C) WT and D) Keap1-disrupted MEF. Effects of LY294002 pre-treatment on WA-mediated Ho-1 induction in E) WT and F) Keap1-disrupted MEF. Gapdh was used as the normalization control for RT-PCR. All values are mean ± SEM (n=3). *p<0.05.