Abstract
Materials and methods Here we use in vitro differentiation of pluripotent mouse embryonic stem (ES) cells to macrophages as a model to identify molecular mechanisms and dynamics of transcriptional programs that define the individual cellular identities. We analyze the chromatin landscape and gene expression of the following five successive cell types that are purified by FACS according to their specific surface marker profiles: mesoderm, hemangioblast, hemogenic endothelium, hematopoietic precursors and macrophages. Genome-wide transcriptional activity of the individual cell populations was assessed by RNA sequencing and the distribution of histone modifications associated with either active or repressed regulatory elements was investigated by ChIP sequencing. Moreover, general DNA accessibility, i.e. open chromatin structure, was detected by DNase 1 digestion and subsequent genomewide sequencing.
Highlights
Cellular identities in multicellular organisms are defined by their individual gene expression programs
Genome-wide transcriptional activity of the individual cell populations was assessed by RNA sequencing and the distribution of histone modifications associated with either active or repressed regulatory elements was investigated by ChIP sequencing
We found a strong correlation between the levels of gene expression and histone modifications
Summary
Nadine Obier1*, Mahalingam S Vijayabaskar, Stella Pearson, Maarten Hoogenkamp, Monika Lichtinger, Pierre Cauchy, David Westhead, Valerie Kouskoff, Georges Lacaud, Bertie Gottgens, Constanze Bonifer. From Epigenetics and Chromatin: Interactions and processes Boston, MA, USA. From Epigenetics and Chromatin: Interactions and processes Boston, MA, USA. 11-13 March 2013
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