Analysis of EMCN+ bone marrow cells in Stab2-iCreF3tg/wt Ctnnb1(Ex3)fl/wt mice

Endothelial-derived Wnt/Ctnnb1-signaling is an important angiocrine regulator. Ctnnb1 gain-of-function (GOF) in sinusoidal endothelial cells was generated via Stab2-iCreF3tg/wt Ctnnb1(Ex3)fl/wt mice to analyze the effects of endothelial β-catenin signaling on bone marrow function. We used microarrays to detail the global programme of gene expression in bone marrow EMCN+ cells with β-catenin (Ctnnb1) gain-of-function compared to Wildtype. Stab2-iCreF3tg/wt Ctnnb1(Ex3)fl/wt and Ctnnb1(Ex3)fl/wt (Stab2-iCreF3 neg) mice were sacrificed at the age of three months, lower limb bones were flushed, digested with 0.7mg/ml of collagenase A (Sigma Aldrich). EMCN+ cells were extracted using EMCN antibody coated magnetic beads. Thereafter, RNA was isolated and prepared for microarray analysis.