Figure 5. Conservative ARE analysis and luciferase reporter analysis of the LAMC1 gene. (A) Comparative transcription factor binding site analysis of the LAMC1 gene across different mammalian species shows highly-conserved NRF2 AREs. (B) Left, siNRF2-C27 and siGFP-C5 cells subjected to ChIP analysis with anti-Nrf2. Right, the relative ability of NRF2 to bind to the ARE site (value of NRF2 in siGFP-C5 cells set at 1; PCR reactions were not saturated; results are from at least 3 separate experiments; HO-1 served as positive control; GAPDH served as negative control). (C) tBHQ increased 151-bp LAMC1 promoter (sequence at left; red indicates ARE sequences)–luciferase activity in MCF7 cells. The plasmid pGL3-LAMC1-151bp was transfected into MCF7 cells in combination with pRL-TK for 24 h. Dual luciferase activity was measured after treatment with tBHQ (20 μM) for 6 h. Control, DMSO treatment for the same plasmid was set at 1 (mean ± SD, n=3; **p <0.01).