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. 2009 Sep 15;18(1):188–197. doi: 10.1038/mt.2009.219

Figure 3.

Figure 3

Mechanism of JG-ODN interacting with HIF-1α and HIF-2α. (a) The results of pull-down assay A DNA gel shows the fluorescent-labeled JG244, and T40214 alone (lanes 1 and 8); GST plus JG243 or JG244 pull-down, showing no interaction between GST and JG-ODNs (lanes 3 and 5); and the pull-down of GST-HIF-1 plus JG243, JG244, JG249, ns-ODN, or T40214, respectively (lanes, 2, 4, and 6-8), showing JG243- and JG244-specific binding with HIF-1 protein but T40214, JG249, and ns-ODN do not. GST-HIF-1 contains the residues from 531 to 826 of HIF-1α. (b) Modeling of JG244 binding within C-terminal of HIF-1α. (c) HIF-1α and HIF-2α levels were determined in lysates from hypoxic untreated MDA-MB-468 breast cancer cells (control) and cells treated with JG243 or JG244 in the presence (+) or absence (−) of MG132. (d) HIF-1α and HIF-2α levels were determined in lysates from normoxia untreated MDA-MB-468 breast cancer cells (control) and cells treated with JG243 or JG244 in the presence (+) or absence (−) of CoCl2. GST, glutathione S-transferase; HIF-1, hypoxia-inducible factor-1; ns ODN, nonspecific oligodeoxynucleotide.