Figure 4.

Antioxidant and xenobiotic detoxification genes are validated as Nrf2 direct targets. (A) and (B) The panels represent ChIP–PCR analysis and densitometry quantification for selected known Nrf2 target gene-binding sites—Txnrd1_P1 and its novel binding site Txnrd1_P2, novel binding sites in Gsta4, three Gstm1-binding sites (Gstm1_P1, Gstm1_P2, Gstm1_P3), Gstm3, Srxn1, Ephx1 and Als2. (C) qRT-PCR mRNA expression analysis in MEFs. (D) Immunoblot analysis for Txnrd1, Gsta4, Gstm1, Srxn1 and Als2 in MEFs. β-Actin was used as the loading control. (E) qRT-PCR for mRNA expression in lung lysates from WT and Nrf2^−/− Air and CS exposed mice, corroborating with the in vitro Nrf2 targets. *P-value < 0.01, as analyzed by ANOVA analysis.