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. 2014 Dec 9;290(5):3057–3068. doi: 10.1074/jbc.M114.621995

FIGURE 7.

FIGURE 7.

Ref-1 associates with NRF2 binding sites on the HMOX-1 promoter and inhibition of Ref-1 diminishes this binding. A, schematic of the NRF2 binding sites on the HMOX-1 promoter (30). B, ChIP assay to show Ref-1 is part of a complex on the HMOX-1 promoter and treatment with the Ref-1 inhibitor can significantly diminish the amount of Ref-1 associated with the HMOX-1 promoter. C, ChIP assay with anti-NRF2 antibody demonstrating that there is more NRF2 bound to the chromatin following Ref-1 inhibition. However, RN7–58 does not increase the amount of NRF2 bound to the HMOX-1 promoter. D and E, Ref-1 siRNA#1 and pooled Ref-1 siRNA#3 and #4 decrease the amount of NRF2 bound to the HMOX1 promoter. MCM5 is included as a negative control to demonstrate that the NRF2-Ref-1 pulldown is not due to nonspecific chromatin binding. IgG and NTC (no template control) serve as controls for the ChIP assay.