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We developed Microarray-before-sequencing (Mab-seq) for use in ChIP experiments with both transcription factor binding and chromatin modification assays.
Gene expression is coordinated by complex regulatory mechanisms that involve transcription factor binding and chromatin modification including DNA methylation.
The differential functions of individual histone variants are characteristic of various processes, such as nucleosome stability, protein binding, and chromatin modification.
Marbach et al. [ 8] furthermore suggest that incorporating additional information such as transcription-factor binding and chromatin modification data or promoter sequences might improve the accuracy of prediction.
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In addition, distal regulatory elements, such as enhancers (defined by DNase I hypersensitivity, transcription factor binding, and chromatin modifications), show differential methylation across tissues (Hon et al., 2013; Ziller et al., 2013).
For mouse embryonic cell CTCF binding sites and chromatin modification marks, we used data by Shen et al. [ 19].
To examine the effects of c-Myc levels on individual genes we analyzed H3-K4 and H3-K27 trimethylation within the subset of Myc associated genes linked to PcG binding, differentiation, pluripotency, and chromatin modification.
We employed a custom-designed oligonucleotide tiling array of 815 promoter regions including promoters of genes that were bound by both Myc and Polycomb proteins as well as other Myc target genes and genes bound by Polycomb but not Myc, selecting genes that are predominantly linked to PcG binding, differentiation, pluripotency, and chromatin modification (Table S4).
For exploring the targets of epigenetic regulation, genome-wide analyses of binding sites and chromatin modifications have been performed in ES cells.
The analysis was performed against both available ChIP types (TF or DNA-binding proteins and chromatin modifications) including both ChIP-chip and ChIP-seq datasets.
Importantly, the applied elongation distance, which corresponds generally to the fragmented chromatin length, is important for proper assignment of a factor binding site or chromatin modification.
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