Histone mark distribution around the Itga2b promoter in EC, ES cells, HPC and mature megakaryocytes.
(A) ChIP was performed using antibodies against H3K9ac, H3K4me3, H3K9me3, H4K8ac and H3K27me3 histone marks. ChiP material were analyzed by Q-PCR and levels of enrichment determined against the control IgG ChIP. (B) Temporal exclusion of the H3K9me3 modification with H3K4me3 and H3K9ac. Sequential ChIP experiments were performed with first a H3K4me3 antibody followed by reChIP using either a H3K9ac or H3K9me3 antibody. The pull down of H3K4me3-associated chromatin was verified by measure of enrichments against the initial input. Maximal enrichment, between +208 and +364 bp from the ATG is represented normalised to the upstream region −1048 to −846 bp (right panel). Equal amount of H3K4me3 ChIP chromatin material were used as inputs for ReChIP experiments with H3K9ac and H3K9me3 specific antibodies. The relative enrichments were determined against the input and normalized against the background as measured across the upstream region −1048 to −846 bp from the ATG. All profiles are representative of 3 independent experiments. Error bars reflect standard error or the mean (SEM).