Analysis of terminal erythroid progenitor populations PIII and PIV in Stab2-iCreF3tg/wt Ctnnb1(Ex3)fl/wt mice

Stab2-iCreF3tg/wt Ctnnb1(Ex3)fl/wt become anemic by the age of three months. We utilzed this model to investigated effects of niche Wnt-Signaling alterations on the terminal erythroid progenitor populations PIII (polychromatic erythroblasts) and PIV (orthochromatic erythroblasts and reticulocytes) by microarray transcriptome analysis. Stab2-iCreF3tg/wt Ctnnb1(Ex3)fl/wt and Ctnnb1(Ex3)fl/wt (Stab2-iCreF3 neg) mice were sacrificed at the age of five months, lower limb bones were flushed and immediately stained for FACS sorting. PIII was defined as SinglecellsLiveCD19negCD3negCD4negCD8negCD11bnegGr1negNk1.1negTer119posCD71posCD44midFSCmid , PIV was defined as SinglecellsLiveCD19negCD3negCD4negCD8negCD11bnegGr1negNk1.1negTer119posCD71posCD44lowFSClow. Cells were lysed directly after sorting and stored at -80°C for 7 days. Thereafter, RNA was isolated and prepared for microarrays.