CnAb1 knock down in R1 mESCs differentiated to mesoderm in Ebs

R1 mESCs were trasnfected with siRNAs against CnAb1 or Control (Luciferase) using Lipofectamine. mRNA was extracted 48 hours using Qiagen RNeasy Mini kit using DNAse treatment Microarray was performed after QPCR validation of the downregulation of CnAb1. Cells were transfected with siRNAs: Control, CnAb1 (1) and CnAb1 (2). For the MicroArray analysis we used 3 control, 3 CnAb1 (1), and 2 CnAb1 (2) samples.

本研究使用Lipofectamine转染试剂，将靶向CnAb1的小干扰RNA（siRNAs）与阴性对照（靶向荧光素酶（Luciferase）的siRNAs）转染至R1小鼠胚胎干细胞（mESCs）。转染48小时后，采用Qiagen RNeasy Mini试剂盒并结合DNase处理提取总mRNA。在通过定量聚合酶链式反应（QPCR）验证CnAb1的基因下调效果后，开展微阵列芯片（Microarray）分析。本次实验共设置三组转染样本：阴性对照组、CnAb1(1)组与CnAb1(2)组；其中用于微阵列芯片分析的样本包括3份阴性对照样本、3份CnAb1(1)组样本以及2份CnAb1(2)组样本。