Fig. 1.

Generation of two murine lines with GFP inserted into the Gata2 locus. (A) Strategy of GFP knock-ins into exon IS or exon II of the Gata2 gene. The WT Gata2 gene (w) has two alternative untranslated first exons (IS and IG). The GFP gene with a polyadenylation signal sequence (pA) and Neo cassette were integrated into exon IS (ISĜFP:neo) or 5′ to the translational start site in exon II (GFP:neo). Neo cassettes were removed by Cre/loxP-mediated site-specific recombination (ISĜFP, GFP). (B) BglII plus NotI, BglII digested genomic DNA from each mouse line was hybridized with radiolabeled exon III probe as indicated in A. (C) Fluorescent images of EIS-KI and EII-KI whole-mount embryos at E11.5. Green fluorescence was observed only in the midbrain (mb) of EIS-KI embryos whereas, in contrast, EII-KI embryos expressed GFP in the placenta (pl), mesonephros (me), midbrain (mb), and the sensory organs (n, nose; e, ear). (D) Mononuclear cells recovered from EII-KI and EIS-KI bone marrow were stained with PE-conjugated antibodies for hematopoietic lineage markers and analyzed by flow cytometry. The percentages of each quadrangle are shown.