ChIP-seq analysis to understand the mechanistic role of NCoR1 as a tolerogenic program suppressor in dendritic cells

We found in our study that NCoR1 knock down dendritic cells (DCs) develop tolerogenic behavior upon activation and these cells have the potential to modulate T helper cell differentiation toward Treg phenotype. Here to understand the mechanism of this process we performed NCoR1 ChIP-seq in wild type DCs before and after CpG ligand activation and transcriptome analysis of NCoR1 KD DCs to identify the direct and indirect target genes. We also predicted PU.1 as the recruting factor for NCoR1 in DCs based on motif enrichment and later on validated that by performing PU.1 ChIP-seq in control and NCoR1 KD DCs. Overall design: Mechanistic role of NCoR1 co-repressor in dendritic cell function

本研究发现，核受体辅阻遏蛋白1（NCoR1）敲低的树突状细胞（DCs）在激活后会呈现致耐受表型，且这类细胞具备调控辅助性T细胞向调节性T细胞（Treg）表型分化的能力。为阐明该过程的分子机制，我们对野生型DCs在CpG配体激活前后开展了NCoR1染色质免疫沉淀测序（ChIP-seq）分析，并对NCoR1敲低DCs进行转录组测序，以鉴定该过程的直接与间接靶基因。此外，基于基序富集分析，我们预测PU.1是DCs中招募NCoR1的转录因子，并通过在对照组与NCoR1敲低DCs中开展PU.1 ChIP-seq实验验证了这一预测。总体实验设计：探究核受体辅阻遏蛋白1（NCoR1）作为辅阻遏因子在树突状细胞功能中的调控机制。