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. 2007 Jan 23;109(10):4181–4190. doi: 10.1182/blood-2005-05-022004

Figure 5.

Figure 5

A flow cytometric analysis of Gfi-1+/− and Gfi-1+/+ EML cells. Gfi-1+/− and Gfi-1+/+ EML cells were stained and analyzed using a FACSvantage flow cytometer and CellQuest software. (A) Cells (1.5 × 105) were then labeled with conjugated antibodies for 20 minutes at 4°C. All antibodies (B220, Sca-1, CD34, c-kit, Mac-1, Gr-1, and Ter119) used for flow cytometry including matched isotype controls were conjugated to phycoerythrin (PE) and purchased from eBiosciences. Error bars indicate SEM. (B) Wright-Giemsa staining of EML+/− and EML+/+ cells following terminal neutrophil maturation. Myeloid differentiation of EML cells was conducted in IMDM medium containing 20% horse serum, IL-3, SCF, and 10 μM ATRA for 3 days. The cells were then transferred to IMDM medium containing 20% horse serum and GM-CSF. Within a week, GM-CSF–dependent EPRO cells emerged. These cells were terminally differentiated by the addition of 10 μM ATRA for 3 days. Cells were cytospun on the days indicated and subjected to Wright-Giemsa staining.