Figure 7

Assessment of the effect of NRF-2 binding elements on the formation of protein/DNA complex. The EMSA analysis was conducted as described in the Methods section. The double-stranded oligonucleotide probe (-311/-282) encompassing both NRF-2(A) and NRF-2(B) binding sites is shown on top. The arrow indicates free probe, and the arrowheads indicate three shifted bands found in the bottom panel. Lane 1 represents the biotin-labeled probe alone. Lane 2 represents the hot probe incubated with nuclear extracts. Lanes 3 and 4 represent the hot probe incubated with nuclear extracts in the presence of either 1- or 10-fold molar excess of cold wild-type probe, respectively. Lanes 5 and 6 represent the hot probe incubated with nuclear extracts in the presence of either 1- or 10-fold molar excess of unlabelled probe with mutation at the NRF-2(B) site, respectively. Lanes 7 and 8 are the same as lanes 5 and 6 except a cold probe with mutation at the NRF-2(A) site was used.