Chromatin Immunoprecipitation of HIF-α in Breast Tumor Cells Using Wild Type and Loss of Function Models

Methods Mol Biol. 2018:1742:67-79. doi: 10.1007/978-1-4939-7665-2_7.

Abstract

Chromatin immunoprecipitation (ChIP) is a powerful method to determine whether a protein of interest binds to specific regulatory elements of the genome. Herein, we outline protocols optimized to detect binding of Hypoxia-Inducible Factor (HIF)-1α or HIF-2α to putative hypoxia response elements (HREs) within HIF target genes expressed in breast tumor epithelial cells.

Keywords: Breast cancer; Chromatin; Cross-linking; Gene deletion; Hypoxia; Hypoxia response element (HRE); Hypoxia-Inducible Factor (HIF); Immunoprecipitation; Input DNA; Polyomavirus middle T (PyMT).

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Aryl Hydrocarbon Receptor Nuclear Translocator / genetics
  • Aryl Hydrocarbon Receptor Nuclear Translocator / metabolism*
  • Binding Sites
  • Breast Neoplasms / genetics*
  • Breast Neoplasms / metabolism
  • Chromatin Immunoprecipitation / methods*
  • DNA / chemistry
  • DNA / metabolism*
  • Female
  • Gene Knockout Techniques
  • Gene Regulatory Networks
  • Humans
  • Hypoxia-Inducible Factor 1, alpha Subunit / genetics
  • Hypoxia-Inducible Factor 1, alpha Subunit / metabolism*
  • MCF-7 Cells
  • Mice
  • Mutation

Substances

  • ARNT protein, human
  • HIF1A protein, human
  • Hypoxia-Inducible Factor 1, alpha Subunit
  • Aryl Hydrocarbon Receptor Nuclear Translocator
  • DNA