Abstract
Epigenetics plays a key role in gene expression control. Histone modifications including acetylation/ deacetylation or methylation/demethylation are major epigenetic mechanisms known to regulate epithelial–mesenchymal transition (EMT)-associated gene expression during hypoxia-induced cancer metastasis. Chromatin immunoprecipitation (ChIP) assay is a powerful tool for investigation of histone modification patterns of genes of interest. In this chapter, we describe a protocol that uses chromatin immunoprecipitation (ChIP) to analyze the epigenetic regulation of EMT marker genes by deacetylation of acetylated Histone 3 Lys 4 (H3K4Ac) under hypoxia in a head and neck cancer cell line FaDu cells. Not only a method of ChIP coupled by real-time quantitative PCR but also the detailed conditions are provided based on our previously published studies.
Original language | English |
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Pages (from-to) | 23-29 |
Number of pages | 7 |
Journal | Methods in Molecular Biology |
Volume | 1436 |
DOIs | |
State | Published - 2016 |
Externally published | Yes |
Bibliographical note
Publisher Copyright:© Springer Science+Business Media New York 2016.
Keywords
- Chromatin immunoprecipitation
- Epithelial–mesenchymal transition
- HDAC3
- Histone modification
- Hypoxia
- Real-time qPCR