E47-deficient HSCs show efficient myeloid differentiation under pathogen-free conditions and after stimulation with LPS. (A-B) Flk2⁻ LSKs from E47 HET or KO mice were cultured at 300 cells per well in 96-well plates for 72 hours in the presence or absence of 10 μg/mL LPS. Cells were harvested, counted, and stained with lineage specific antibodies. The number and frequency of CD11b⁺ myeloid cells were measured (n = 9 wells). **P < .05. ns indicates not significant. The data are representative of 2 independent experiments. (C) The expression of CD14 on flk2⁻ LSKs and flk2⁺ LSKs from WT and KO littermates was examined by flow cytometric analysis. The data are representative of 2 independent experiments. (D) flk2⁻ LSKs from E47 WT or KO mice were cultured in MethoCult at 1, 2, and 5 cells per well in 96-well plates for 7 days in the presence or absence of 10 μg/mL LPS. Wells with colony-forming unit-granulocyte, macrophage colonies were scored positive. The frequency of colony-forming cells was calculated according to Poisson statistics (n = 48 wells per group). The data are representative of 2 independent experiments.