RNA-sequencing analysis of colon RNA from mice with compound mutations of XBP1, caspase-8, TNFR1 and MLKL.

To purpose of this study was to assess how intestinal epithelial cell (IEC)-specific ablation of XBP1 or caspase-8 alone or in combination affected mucosal immune homeostastis and inflammation, and the role of TNFR1 and MLKL in the associated phenotypes. First, expression of colonic RNA from mice with combined IEC-specific XBP1 and caspase-8 ablation (Xbp1IEC-KO Casp8IEC-KO) was compared with Casp8IEC-KO, Xbp1IEC-KO, and wild-type mice. In addition, Xbp1IEC-KO Casp8IEC-KO Mlkl-/- and Xbp1IEC-KO Casp8IEC-KO Tnfr1IEC-KO mice were compared with Xbp1IEC-KO Casp8IEC-KO, Casp8IEC-KO, Xbp1IEC-KO as well as wild-type control mice. For each mouse line 5 individual mice were analysed. RNAs from the colons of control, Xbp1IEC-KO, Casp8IEC-KO, Xbp1IEC-KO Casp8IEC-KO, Xbp1IEC-KO Casp8IEC-KO, Xbp1IEC-KO Casp8IEC-KO Tnfr1IEC-KO and Xbp1IEC-KO Casp8IEC-KO Mlkl-/- mice were isolated. Then RNA-sequencing-assisted gene expression analysis were performed from these isolated RNAs.本研究旨在探究单独或联合敲除肠上皮细胞(Intestinal Epithelial Cell, IEC)特异性的XBP1或半胱天冬酶-8(caspase-8)，对黏膜免疫稳态与炎症的影响，以及肿瘤坏死因子受体1(TNFR1)和混合谱系激酶结构域样蛋白(MLKL)在相关表型中的作用。首先，将同时特异性敲除IEC中XBP1和caspase-8的小鼠（Xbp1IEC-KO Casp8IEC-KO）的结肠RNA表达水平，分别与Casp8IEC-KO小鼠、Xbp1IEC-KO小鼠以及野生型小鼠进行比较。此外，将Xbp1IEC-KO Casp8IEC-KO Mlkl-/-小鼠及Xbp1IEC-KO Casp8IEC-KO Tnfr1IEC-KO小鼠，与Xbp1IEC-KO Casp8IEC-KO、Casp8IEC-KO、Xbp1IEC-KO及野生型对照小鼠进行对比。每种小鼠品系均选取5只个体进行分析。分别提取对照、Xbp1IEC-KO、Casp8IEC-KO、Xbp1IEC-KO Casp8IEC-KO、Xbp1IEC-KO Casp8IEC-KO、Xbp1IEC-KO Casp8IEC-KO Tnfr1IEC-KO以及Xbp1IEC-KO Casp8IEC-KO Mlkl-/-小鼠的结肠RNA。随后对提取的RNA开展RNA测序(RNA-sequencing)辅助的基因表达分析。