ChIP-seq and RNA-seq analysis of histone Kcr in the embryonic forebrain and neural stem/progenitor cells.. ChIP-seq and RNA-seq analysis of histone Kcr in the embryonic forebrain and neural stem/progenitor cells.

To gain a deeper insight into how histone lysine crotonylation regulates embryonic neural stem/progenitor cells (NSPCs) proliferation and differentiation, ChIP-seq and RNA-seq were performed to analyze the genome-wide changes of histone Kcr and transcriptiome changes stimulated with crotonate and MS-275 in E13.5 NSPCs. Overall design: Total RNA was extracted from E13.5 NSPCs before and after crotonate and MS-275 stimulation and generated by deep sequencing, in four biological replicates, using Illumina novaseq 6000. ChIP was excuted using pan-Kcr and Pan-Kac antibody with E13.5 NSPCs before and after crotonate and MS-275 stimulation.

为深入解析组蛋白赖氨酸巴豆酰化（histone lysine crotonylation）对胚胎神经干细胞/前体细胞（embryonic neural stem/progenitor cells, NSPCs）增殖与分化的调控机制，本研究采用染色质免疫共沉淀测序（ChIP-seq）与RNA测序（RNA-seq）技术，分析了巴豆酸盐与MS-275处理的E13.5 NSPCs的全基因组组蛋白Kcr修饰变化及转录组改变。总体实验设计如下：总RNA提取自巴豆酸盐与MS-275处理前后的E13.5 NSPCs，依托Illumina NovaSeq 6000平台开展深度测序，设置4个生物学重复；染色质免疫共沉淀实验以经巴豆酸盐与MS-275处理前后的E13.5 NSPCs为实验材料，使用泛巴豆酰化抗体（pan-Kcr）与泛乙酰化抗体（Pan-Kac）完成检测。