Figure 3. ET-1/ET_AR stimulates the recruitment of β-arr1, HIF-1α and p300 on the promoter of ET-1 and VEGF target genes.

A. HEY cells were treated with ET-1 for indicated times and the binding of HIF-1α or β-arr1, to ET-1 promoter region was measured by ChIP assays followed by PCR. HEY cells were treated with ET-1 and/or MAC for 2 h and the binding of HIF-1α or β-arr1, or p300, or acetylated histone 3 (AcH3) to ET-1 promoter B. or to VEGF promoter C. region was measured by ChIP assays followed by PCR. D. HEY cells were transfected with SCR or si- β-arr1 and treated with ET-1 for 2 h and the binding of β-arr1, HIF-1α, AcH3, and p300 to ET-1 promoter region was measured by ChIP assay followed by PCR. Non-specific IgG was used as the irrilevant antibody (IRR) for all ChIP reactions. The input DNA lane represents one- twentieth of the precleared chromatin used in each ChIP reaction. E. qRT-PCR analysis of ET-1 and VEGF expression in HEY cells transfected with SCR or si- β-arr1 and stimulated with ET-1 and/or MAC. Bars are means ±S.D from three independent experiments each performed in triplicate; *, p < 0.002 versus CTR transfected cells; **, p < 0.001 versus ET-1.