Yap/Taz negatively regulate bone angiogenesis and osteogenesis

In long bone, the metaphyseal region is less hypoxic than the bone marrow (bone shaft). To study the regional differences of vasculatures, we isolated endothelial cells (ECs) expressing membrane-anchored mTomato protein and nuclear H2B-GFP from Cdh5-mT/nG transgenic reporter femoral metaphysis and diaphyseal bone marrow cavity by fluorescence-activated cell sorting (FACS) at high purity for RNA sequencing (RNA-seq). Next, to study the Hippo signaling in bone ECs, we performed RNA-seq analysis of freshly isolated bone ECs from Cdh5Cre-ERT2-positive Yap1/Taz double floxed or Cdh5Cre-ERT2-positive Lats2 floxed allele and their Cre-negative corresponding littermate controls. Control and Cdh5(PAC)CreERT2-Yap1; Wwtr1 mutant bone endothelial cells were isolated by using FACS. RNA was isolated from control and EC-Yap1; Wwtr1 mutant endothelial cells. Bone endothelial cells (Cdh5 mT/nG+) from metaphyseal and marrow transcripts were analyzed. We studied gene expression changes in absence of Yap1/Taz and Lats2 in bone endothelial cells.