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. Author manuscript; available in PMC: 2014 Jun 9.
Published in final edited form as: FASEB J. 2007 Apr 5;21(10):2613–2621. doi: 10.1096/fj.06-8004com

Figure 4.

Figure 4

VASP promoter analysis and HIF-1α binding analysis. a) Map of cloned VASP construct utilized here. Relative positions of each clone are annotated, as well as the transcription start site (TSS). b) HMEC-1 or HeLa cells, as indicated, were transfected with human VASP pGL3 reporter vector and exposed to normoxia or normobaric hypoxia (1% O2, 99% N2) for 24 hours post transfection (n=4). Results are presented as relative luciferase activity±sem, where * indicates p<0.025). c) ChIP assay analysis performed with HIF-1α antibody in HMEC-1 and T84 exposed to 24h normobaric hypoxia. d) Site-directed mutagenesis of HIF-1α binding site located at position - 522 to – 519 of pGL3 reporter construct shows attenuated VASP downregulation during hypoxia (n=4). Results are presented as relative luciferase activity±sem, where * indicates p<0.025). Also shown is a positive control plasmid encoding the HRE from the erythropoietin gene (EPO HRE).