Chip-seq of H3 in murine embryonic stem cells, myotubes and pro-B cells.

Nucleosome remodeling results in loss of histone occupancy. To gain insight into variations in H3 occupancy in different murine cell types, chromatin immunoprecipitation coupled with massive parallel sequencing (ChIP-seq) was performed to determine genome wide occupancy of H3 in ES cells, myotubes and pro-B cells. Overall design: DNA was enriched by chromatin immunoprecipitation (ChIP) and analyzed by Solexa sequencing. ChIP was performed using an antibody against total H3 in murine ES cells in normal culture conditions, murine ES cells treated with the TGF-beta inhibitor (SB431542, 10uM) for 24 hours, myotubes differentiated for 48 hours and pro-B cells under normal culture conditions.

核小体重塑会导致组蛋白占据率下降。为探究不同小鼠细胞类型中H3占据率的差异，本研究采用染色质免疫共沉淀结合大规模平行测序（chromatin immunoprecipitation coupled with massive parallel sequencing，ChIP-seq）技术，检测了胚胎干细胞（Embryonic Stem Cell, ES细胞）、肌管细胞（myotubes）以及前B细胞（pro-B cells）的全基因组H3占据情况。总体实验设计：通过染色质免疫共沉淀（ChIP）富集DNA片段，并采用Solexa测序进行分析。本次ChIP实验使用针对总H3的抗体，所使用的样本包括：常规培养条件下的小鼠ES细胞、经转化生长因子β（TGF-β）抑制剂SB431542（10μM）处理24小时的小鼠ES细胞、诱导分化48小时的肌管细胞，以及常规培养条件下的前B细胞。