Role of Sox9 in Yap activated hepatocytes

Hepatocyte specific TBG promoter-driven Cre,GFP and YAP S127A expression was achieved by retro orbital injection of AAV. RNA-seq was performed for hepatocytes isolated by FACS. For YAP S127A overexpression groups, GFP+ control and YapS127A expressing hepatocytes were isolated by FACS 3 weeks after injection. For Mst1, Mst2, Sox9 conditional knockout; Rosa26-lsl-TdTomato groups, GFP+ control and TdTomato expressing mutant hepatocytes were isolated by FACS 4 weeks after injection.

通过经眶后注射腺相关病毒（AAV），成功实现肝细胞特异性TBG启动子驱动的Cre、GFP与YAP S127A的表达。对经荧光激活细胞分选（FACS）分离得到的肝细胞开展RNA测序（RNA-seq）。在YAP S127A过表达实验组中，于病毒注射后3周通过FACS分选出GFP阳性对照肝细胞与表达YapS127A的肝细胞。针对Mst1、Mst2、Sox9条件性敲除且携带Rosa26-lsl-TdTomato的实验组，则于注射后4周通过FACS分选出GFP阳性对照肝细胞与表达TdTomato的突变肝细胞。