RNA-seq profiling of foetal-derived Langerhans cells from wildtype and Suz12 knockout mice

Macrophages are at the forefront of immune responses and their transcriptional programs are modified by their tissue environment and in response to immunological challenge. Post-translational modifications of histones, such as histone H3 lysine 27 tri-methylation (H3K27me3) by the polycomb repressive complex 2 (PRC2), are tightly associated with epigenetic regulation of gene expression. To explore whether H3K27me3 is involved in either the establishment or function of the mononuclear phagocyte system, we selectively deleted the SUZ12 gene in one-week old mice, a core component of PRC2. Overall design: SMART-Seq RNA libraries were generated from 5000 purified Langerhans cells from the skin of one week old wild mice (2 biological replicates) or Suz12cKO (3 biological replicates). Samples were sequenced with an Illumina NextSeq 500 sequencing system, producing between 22-34 million paired-end 75 bp reads per sample.

巨噬细胞（macrophages）是免疫应答的前沿防线，其转录程序可受组织微环境调控，并可响应免疫挑战而发生重塑。组蛋白的翻译后修饰，例如由多梳抑制复合体2（polycomb repressive complex 2, PRC2）介导的组蛋白H3赖氨酸27三甲基化（histone H3 lysine 27 tri-methylation, H3K27me3），与基因表达的表观遗传调控紧密相关。为探究H3K27me3是否参与单核吞噬细胞系统（mononuclear phagocyte system）的建立或功能维持，我们在1周龄小鼠中特异性敲除了PRC2的核心组分SUZ12基因。
实验设计概述：从1周龄野生型小鼠（2个生物学重复）或Suz12条件性敲除小鼠（Suz12cKO，3个生物学重复）的皮肤中纯化分离5000个朗格汉斯细胞（Langerhans cells），以此构建SMART-Seq RNA文库；随后采用Illumina NextSeq 500测序系统对样本进行测序，每个样本可产生2200万至3400万条75bp双端测序读段。