Effect of BAF disruption on T-bet binding in activated CD8+ T cells [ChIP-seq]

To investigate the role of cBAF in regulation of Tbet binding, polyclonal naive CD8 T cells were stimulated for 48h with anti-CD3, CD28, and hIL-2. Cells were then treated with DMSO, ACBI1, or BRM014 for 6 hours, and treated with recombinant mouse IL-12 for the last two hours of inhibitor treatment. ChIP-seq against T-bet was then performed. Overall design: T-bet ChIP-seq on WT CD8+ T cells activated with anti-CD3, anti-CD28, and hIL-2 for 48h in vitro with or without IL-12, ACBI-1, or BRM014.

为探究cBAF对T-bet结合的调控作用，本研究将多克隆初始CD8 T细胞采用抗CD3、抗CD28及人源白细胞介素2（hIL-2）体外刺激48小时。随后将细胞分别用二甲基亚砜（DMSO）、ACBI1或BRM014处理6小时，并在抑制剂处理的最后2小时加入重组小鼠白细胞介素12（IL-12）。之后开展针对T-bet的染色质免疫沉淀测序（ChIP-seq）。
实验整体设计：对经抗CD3、抗CD28及hIL-2体外激活48小时的野生型（WT，Wild Type）CD8+ T细胞开展T-bet ChIP-seq实验，实验分组包含或不添加IL-12、ACBI1或BRM014处理。