E2A Antagonizes PU.1 Activity through Inhibition of DNA Binding

Figure 4

E47 inhibits myeloid differentiation of hematopoietic cells. (a) PUER cells were superinfected with MigR1 or MigR1-E47 virus and pure GFP expressing clones generated. Cell lines were either undifferentiated (no OHT) or differentiated (100 nM OHT) for 8 d. Immunohistochemistry of adherent PUER cell lines induced to differentiate with 100 nM OHT for 8 d. Cells were stained either with the macrophage differentiation marker F4/80 or with normal rat serum (NRS). Magnification is 400x. (b) F4/80 surface expression as determined by flow cytometry of independent PUER GFP and PUER E47 lines with no addition of OHT and after 8 d of OHT treatment. (c) Flow cytometry analysis of lineage minus hematopoietic progenitors infected with control MigR1 virus or MigR1-E47 virus. Cells were cultured for 12 d on OP9 stromal cell layer in the presence of 1 ng/mL IL-7 and 5 ng/mL Flt3L. Differentiation was analyzed by staining cells with anti-CD11b (myeloid) and anti-CD19 (B cell). Percentages of total cells positive for CD11b and CD19 expression are shown in the upper right corner of FACs plots.

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