Expression data from differentiating Flk1- and Flk1+ ES cells expressing Snail during Wnt inhibition

ES cells differentiated in the presence of the Wnt inhibitor DKK1 fail to express the transcription factor Snail and undergo EMT or mesoderm differentiation. We generated an ES cell line, A2.snail, that induced Snail expression upon addition of doxycycline addition. Microarrays were used to gain a global picture of Flk1- and Flk1+ cells generated one day after Snail was expressed during Wnt inhibition. A2.snail ES cells, which express Snail upon addition of doxycycline, were differentiated as embryoid bodies in differentiation media and DKK1. Snail-induced cultures uniquely develop a select population of Flk1+ cells. Total RNA was harvested from sorted control (no doxycycline) Flk1- cells and sorted Snail-induced (doxycycline at day 2) Flk1- and Flk1+ cells at day 3 of differentiation.

在Wnt信号通路抑制剂DKK1存在的培养体系中分化的胚胎干细胞（ES cells），无法表达转录因子Snail，且不会发生上皮间质转化（EMT, Epithelial-Mesenchymal Transition）或中胚层分化。我们构建了一株ES细胞系A2.snail，该细胞系在加入多西环素（doxycycline）后可诱导Snail的表达。我们采用基因芯片（Microarrays）技术，获取了Wnt抑制环境下Snail表达1天后产生的胎肝激酶1阴性（Flk1-）与胎肝激酶1阳性（Flk1+）细胞的全局基因表达谱。我们将可在加入多西环素后表达Snail的A2.snail ES细胞置于添加了DKK1的分化培养基中，以拟胚体（embryoid bodies）的形式进行分化。经Snail诱导的培养体系可特异性生成特定的Flk1阳性细胞群。在分化第3天，我们从流式分选获得的对照组（未添加多西环素）Flk1阴性细胞，以及经Snail诱导（第2天添加多西环素）的流式分选Flk1阴性与Flk1阳性细胞中提取了总RNA。