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. 2009 May 22;284(21):14001–14010. doi: 10.1074/jbc.M809320200

FIGURE 3.

FIGURE 3.

A, effect of echinomycin on RON promoter activity: cells were transfected with pGL3 control vector or –400-bp wild-type RON promoter. 24 h following transfection, cells were treated with varying concentrations of echinomycin for 16 h, and luciferase activity was measured following normalization to protein levels. B, ChIP assay on the chromatin fragments from breast cancer cells using control IgG or HIF-1α antibodies was performed as described under “Experimental Procedures.” C, illustration: RON promoter-luciferase reporter construct depicting the HIF-1α binding site (HRE) and Sp1 binding sites. D, effect of HRE binding site mutation on RON promoter activity: cells were transfected with either pGL3 control vector or –400-bp wild-type RON promoter or HRE binding site mutant RON promoter-luciferase reporter constructs and luciferase activity was measured after 48 h following normalization to protein levels. To determine the effect of HIF-1α expression knockdown on RON promoter activity control or –400-bp wild-type RON promoter activity was analyzed in control shRNA or shRNA HIF-1α-expressing clones.