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. Author manuscript; available in PMC: 2015 Jun 1.
Published in final edited form as: Biochim Biophys Acta. 2014 Apr 2;1839(6):493–505. doi: 10.1016/j.bbagrm.2014.03.014

Figure 4. Role of DNase HS sites in Nrf2-mediated induction of E1b.

Figure 4

Figure 4

A.) Effect of Nrf2 overexpression or tBHQ treatment on transactivation activity of E1b DNase I HS sites in BEAS-2B and A549 cells. For Nrf2 overexpression, cells were co-transfected with E1b promoter reporter constructs containing HS-1 or HS-2 and varying amounts of Nrf2. Luciferase activity was determined 24 h later. For tBHQ induction experiments, cells were transfected with promoter reporter constructs containing HS-1 or HS-2 and treated with varying amounts of tBHQ 18 h later. Luciferase activity was determined 6 h after treatment. KEAP1 down regulates HS-2 enhancer activity in A549 cells (B) and SFN-treated BEAS-2B cells (C). Cells were co-transfected with the KEAP1 expression plasmid and the reporter vectors containing E1b-300 promoter or E1b DNase I HS-2/E1b-300 promoter for 24 h. A549 cells were harvested for luciferase assay while BEAS-2B cells were treated with 2μM SFN for extra 24 h before luciferase assay. All luciferase values represent mean ± SD. *p<0.05 compared to empty vector controls or DMSO control.