Expression of the Nrf2 target genes NQO- 1 and AKR1C1 after t-BHQ treatment. Real-time PCR analysis of the Nrf2 target genes NQO-1 (A, left) and AKRC1 (B, left) in HT29 cells (methylated) and Colo320DM cells (unmethylated). Cells were treated with the Keap1 stimulator t-BHQ for 24 h. Columns, mean (n = 3); bars, SD. *P < 0.05. Real-time PCR analysis of NQO-1 (A, right) and AKRC1 (B, right) in HT29 cells treated with 5-Aza-dC + TSA and then t-BHQ (for 12 h). Keap1 protein levels (C, upper), protein expression of downstream ARE genes (C, upper), and changes in the subcellular distribution of Nrf2 protein (C, lower) after 5-Aza-dC and TSA treatment. Cells were treated with 5-Aza-dC and TSA. Expression levels of each protein were determined by Western blotting. β-actin and histone H1 were used as loading controls.