Fig. 4

Over-expression of EAR-2 in vitro inhibits hematopoietic differentiation while knockdown increases hematopoietic differentiation. Bone marrow transduced with either EAR-2 or empty vector (GFP) was analyzed for a colony formation in methylcellulose medium, (b i and ii) and differentiation ability in suspension culture followed by immunophenotypic analysis using flow cytometry. c Serial colony formation assays of bone marrow ex vivo transduced with EAR-2 shows an increased proliferative capacity ex vivo. Silencing of murine BM with shRNA did not effect (d i) colony number, but greatly increased colony size in conditions favoring (d ii) multilineage and (d iii) erythroid differentiation. (d iv) Silencing of EAR-2 decreased serial colony forming ability of bone marrow. e Gene silencing of EAR-2 depleted bone marrow of undifferentiated cells (the lineage negative fraction) in short-term culture, f such that the cells differentiated into granulocytes