Journal: Proceedings of the National Academy of Sciences of the United States of America
Article Title: Inhibition of vascular endothelial growth factor with a sequence-specific hypoxia response element antagonist
Figure Lengend Snippet: Polyamide 1 binds HRE. ( A ) Storage phosphor autoradiograms from quantitative DNase I footprint titrations of polyamides 1 and 2. The boxed sequence ( Left ) represents the HRE site. For polyamide 1, lanes 1 and 16, A reaction: lane 2, intact DNA; lanes 3–14, DNase I digestion products in the presence of 50 nM, 20 nM, 10 nM, 5 nM, 2 nM, 1 nM, 500 pM, 200 pM, 100 pM, 50 pM, 20 pM, and 10 pM polyamide, respectively; lane 15, DNase I standard. For polyamide 2, lanes 1 and 16, A reaction: lane 2, intact DNA; lanes 3–14, DNase I digestion products in the presence of 1 μM, 300 nM, 100 nM, 30 nM, 10 nM, 3 nM, 1 nM, 300 pM, 100 pM, 30 pM, 10 pM, and 3 pM polyamide, respectively; lane 15, DNase I standard. ( B ) Storage phosphor autoradiogram from EMSA experiment with polyamides 1 and 2.
Article Snippet: Genomic DNA was digested by treatment with DNase I from a DNA Free kit (Ambion, Austin, TX), and DNase I was inactivated with bead-immobilized DNase I inactivation reagent (Ambion).