Chromatin Immunoprecipitation Assay for Transcription Factors

Chromatin was extracted from cells (5 × 10⁶) and ChIP assays were performed as previously described [18 (link)]. The differential binding between proteins and promoter DNA was examined by PCR. The primary Abs used were as follows: HIF-1α (Abcam, UK), β-arr1 (Santa Cruz Biotechnology CA, USA), p300 (Santa Cruz Biotechnology CA, USA) and acetyl-Histone 3 (AcH3) (BD Laboratory Transduction, NJ, USA). The primers used were as follows: ET-1 promoter Fw: 5′-CAGCTTGCAAAGGGGAAGCG-3′ and Rev: 5′-TCCGACTTTATTCCAGCCCC-3′; VEGF promoter Fw: 5′-AGGAACAAGGGCCTCTGTCT-3′ and Rev: 5′-CAGTGTGTCCCTCTGACAATG-3′.