RNA-seq Analysis of the gene expression regulated by CID755673 in mouse embryonic stem cells. RNA-seq Analysis of the gene expression regulated by CID755673 in mouse embryonic stem cells

Purpose:The goals of this study is to analyze the candidate genes motified by CID755674 to promote the mouse embryonic stem cell (ESC) self-renewal through RNA-seq approach. Previously, we found that MEK inhbitior PD0323901 (PD) coporates with protein kinase D inhibitor CID755673 (CID) is able to maitain mouse embryonic stem cell (mESC) self-renewal, we named this conditon is "PC". In order to find out why CID755673 can promote mouse embryonic stem cell self-renewal, we performed RNA-sequence in mESCs treated with PD or PC, and tried to identify the key mechanisms under CID Overall design: 46C mouse ESCs cultured in 1 µM PD0325901/serum condition in the presence or absence of CID755673 (named PD vs PC) for 24 hr and then were collected by using Trizol. Two repeat groups were set for each treatment group. These samples were sent to GENERGY BIO (Shanghai, China) company to do deep sequencing by using illumina Hiseq.

研究目的：本研究旨在通过RNA测序（RNA-seq）技术，分析受CID755673调控的候选基因，以阐明其促进小鼠胚胎干细胞（ESC）自我更新的分子机制。此前本团队发现，MEK抑制剂PD0325901（PD）与蛋白激酶D抑制剂CID755673（CID）联合使用，可维持小鼠胚胎干细胞（mESC）的自我更新能力，我们将该培养体系命名为“PC”。为探明CID755673促进小鼠胚胎干细胞自我更新的关键机制，本研究对经PD或PC处理的小鼠胚胎干细胞开展RNA测序，以期解析CID介导的核心调控机制。
实验设计：将46C小鼠胚胎干细胞置于含1 µM PD0325901/血清的培养体系中，分别添加与不添加CID755673（即PD组 vs PC组），培养24小时后采用Trizol法收集样本。每个处理组设置2个生物学重复。所有样本送至中国上海GENERGY BIO公司，采用Illumina Hiseq平台进行深度测序。