Fig. 2.

High-level expression of GFP and GATA-2 mRNA in HSC of E1S-KI mice. (A) Lineage negative (Lin⁻) cells recovered from EIS-KI bone marrow were analyzed by flow cytometry after staining with c-Kit and Sca1 antibodies. Cells were also stained by Hoechst 33342, and blue and red fluorescence were measured (H. Blue and H. Red, respectively) to detect the side population (SP). The percentage of cells in each quadrangle is shown. (B) Cells in each quadrangle of the Lin⁻c-Kit⁺ fraction (Lower Left in A) were compared for levels of GATA-2 mRNA by quantitative RT-PCR (normalized to GAPDH). (C) The morphology of Lin⁻GFP⁺Sca1⁺c-Kit⁺ (G⁺L⁻S⁺K⁺) cells from EIS-KI bone marrow are shown after Wright-Giemsa staining. (Scale bar: 10 μm.) (D) Cell-cycle status of G⁺L⁻S⁺K⁺ and G⁻L⁻S⁺K⁺ cells. GFP⁺ and GFP⁻ cells in Sca1⁺c-Kit⁺ fraction of fixed EIS-KI bone marrow were analyzed for their DNA content by Hoechst 33342 flow cytometry. The percentages of cells in S/G₂/M-phases are shown.