The two histone H2AZ variant, H2AZ.1 and H2AZ.2, differentially regulate PRC1-mediated histone H2A ubiquitinatione

We performed CUT&Tag experiments in mouse embryonic stem cells (mESCs) to investigate the chromatin binding of H2AZ1 and Ring1B. H2AZ1 CUT&Tag was conducted in H2AZ1KIFlag mESCs using an anti-Flag antibody. To assess how knockdown of H2AZ1 and H2AZ2 affects Ring1B occupancy on chromatin, we performed additional CUT&Tag experiments in Ring1BKIFlag mESCs under knockdown conditions. This study aims to elucidate the regulatory interplay between histone variants and Polycomb proteins in the context of pluripotent stem cell chromatin. Overall design: CUT&Tag was performed to profile the chromatin binding of H2AZ1 and Ring1B in mouse embryonic stem cells (mESCs). H2AZ1 binding was assessed in H2AZ1KIFlag mESCs using an anti-Flag antibody. To investigate the effects of H2AZ1 and H2AZ2 knockdown on Ring1B occupancy, CUT&Tag experiments were conducted in Ring1BKIFlag mESCs under knockdown conditions for H2AZ1 and H2AZ2. Each condition included biological replicates. Sequencing libraries were prepared and sequenced as paired-end reads. Both raw data are included in this submission.

我们在小鼠胚胎干细胞（mESCs）中开展CUT&Tag实验，以探究H2AZ1与Ring1B的染色质结合特征。H2AZ1的CUT&Tag实验在携带H2AZ1KIFlag标签的mESCs中进行，采用抗Flag抗体进行靶标富集。为评估H2AZ1与H2AZ2的基因敲降对Ring1B染色质占据情况的影响，我们在携带Ring1BKIFlag标签的mESCs中，于H2AZ1和H2AZ2敲降条件下开展了额外的CUT&Tag实验。
本研究旨在阐明多能干细胞染色质背景下，组蛋白变体与Polycomb蛋白之间的调控互作关系。
整体实验设计：本研究通过CUT&Tag技术对小鼠胚胎干细胞（mESCs）中H2AZ1与Ring1B的染色质结合谱进行分析。H2AZ1的结合情况通过抗Flag抗体在携带H2AZ1KIFlag标签的mESCs中检测。为探究H2AZ1与H2AZ2敲降对Ring1B染色质占据情况的影响，我们在H2AZ1和H2AZ2敲降条件下的携带Ring1BKIFlag标签的mESCs中开展CUT&Tag实验。每组实验均设置生物学重复。测序文库构建完成后，采用双端测序策略进行测序。本次提交包含全部原始测序数据。