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. 2012 Apr 4;32(6):1046–1060. doi: 10.1038/jcbfm.2012.21

Figure 6.

Figure 6

Hypoxia alters DNA binding ability of hypoxia-inducible factor-1α (Hif-1α) and nuclear factor erythroid 2-related factor 2 (Nrf2) through neuroglobin (Ngb)-mediated signaling. Nuclear fraction of proteins was prepared from in-vivo and in-vitro cell lysates and incubated with suitable probes for Hif-1α and Nrf2. Panels (A, B) and corresponding graphs showing representative electrophoretic mobility shift assay (EMSA) for DNA binding ability of Hif-1α and Nrf2 in hippocampus of normoxic and hypoxic rats. Panels (C, D) and corresponding graphs showing representative EMSA for DNA binding ability of Hif-1α and Nrf2 in N2a cells. Nuclear fractions incubated with immobilized primary antibodies for Hif-1α and Nrf2 were considered as negative controls. Graphs denote percentage of DNA binding as determined by optical density of the bands, considering normoxic control values to be 100%. *Denotes P<0.01 when compared with normoxic controls. H, hypoxia; NAC, N-acetyl cysteine; Si, silenced; OE, overexpressed; NC, negative control.