High Yap and Mll1 promote a persistent regenerative cell state induced by Notch signaling and loss of p53

Murine small intestinal crypts of six independent mice were isolated and cultured as 3D-Organoids. Expression of NICD and loss of p53 mutagenesis was induced by 24h treatment with 4OH-tamoxifen in three of the cultures, resulting in three mutant organoid lines. The other three organoid lines were not exposed to tamoxifen and served as controls. Of each organoid line a duplicate was taken resulting in six control and six mutant samples that were compared in the analysis.

从6只独立小鼠体内分离得到小肠隐窝（small intestinal crypts），并将其培养为3D类器官（3D-Organoids）。选取其中3份培养体系，采用4-羟基他莫昔芬（4OH-tamoxifen）处理24小时，以诱导Notch胞内域（NICD）表达并通过诱变实现p53功能丧失，最终构建得到3株突变型类器官系。剩余3株类器官系未接触他莫昔芬，作为对照组。每株类器官系均设置生物学重复样本，最终获得6份对照样本与6份突变样本用于后续分析比对。