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