Zebrafish whole embryos at 36 hpf treated with DMSO or 5uM 11,12-EET

In our previous study, we found zebrafish embryos treated with 5uM 11,12-EET (epoxyeicosatrienoic acid) had increased stem cell marker, runx1, expression in the AGM. EET also induced ectopic runx1 expression in the tail. To systematically study how EET regulates gene expression, we performed microarray analysis on EET-treated embryos. Zebrafish whole embryos were synchronized at fertilization. Embryos were grown at 28 degree overnight. 25 embryos per group were treated with DMSO or 5uM 11,12-EET starting from 24 hpf (hour post fertilization) until 36 hpf at 28 degree. The triplicates were from three different clutches of embryos, and split into DMSO v.s. EET for each clutch. EET vs. DMSO

在本团队此前的研究中，我们发现经5μM 11,12-环氧二十碳三烯酸（epoxyeicosatrienoic acid，简称11,12-EET）处理的斑马鱼胚胎，其主动脉-性腺-中肾区（aorta-gonad-mesonephros，AGM）内干细胞标志物runx1的表达水平显著升高。此外，EET还可诱导斑马鱼尾部出现异位的runx1表达。为系统探究EET对基因表达的调控作用，我们对经EET处理的斑马鱼胚胎开展了基因芯片分析。实验具体流程如下：将斑马鱼全胚胎于受精后进行同步化处理，随后在28℃条件下培养过夜。自受精后24小时（hour post fertilization，hpf）起，每组取25枚胚胎，在28℃条件下分别用二甲基亚砜（DMSO）或5μM 11,12-EET处理至受精后36 hpf。本实验设置3次生物学重复，样本均来自3个独立的胚胎批次，每个批次均单独分为DMSO对照组与EET处理组。EET处理组 vs. DMSO对照组