Characterizing Epigenetic Changes in Endothelial Cells.

Chromatin immunoprecipitation (ChIP) is an antibody-based method used to identify protein-DNA interactions and sites of protein modifications to chromatin in living cells. ChIP is a powerful method for identifying genomic sites at which epigenetic changes occur in cell types of interest because many antibodies have been developed that recognize specific epigenetic modifications of histone tails. This chapter provides detailed ChIP and subsequent polymerase chain reaction (ChIP-PCR) protocols for use in cultured endothelial cells.

BRG1 (Brahma-Related Gene 1) Promotes Endothelial Transcription to Establish Embryonic Capillary Integrity.

Objective: The chromatin remodeling enzyme BRG1 (brahma-related gene 1) transcriptionally regulates target genes important for early blood vessel development and primitive hematopoiesis. However, because deletion in vascular progenitor cells results in lethal anemia by embryonic day 10.5 (E10.

siRNA Screening Identifies the Host Hexokinase 2 (HK2) Gene as an Important Hypoxia-Inducible Transcription Factor 1 (HIF-1) Target Gene in Toxoplasma gondii-Infected Cells.

Unlabelled: Although it is established that oxygen availability regulates cellular metabolism and growth, little is known regarding how intracellular pathogens use host factors to grow at physiological oxygen levels. Therefore, large-scale human small interfering RNA screening was performed to identify host genes important for growth of the intracellular protozoan parasite Toxoplasma gondii at tissue oxygen tensions. Among the genes identified by this screen, we focused on the hexokinase 2 (HK2) gene because its expression is regulated by hypoxia-inducible transcription factor 1 (HIF-1), which is important for Toxoplasma growth.