![Figure 1. Cis-regulatory element requirement for Gata2 expression and hematopoietic progenitor activity in the AGM. (A) Schematic diagram for targeted deletion of the E-box-GATA composite element in murine Gata2 locus. (B) AGM ex vivo culture system. (C) Representative images of E11.5 +9.5+/+,+9.5+/−, and +9.5−/− embryos. (D) Real-time RT-PCR analysis of Gata2 expression levels in E11.5 uncultured AGMs, cultured AGM explants, and cultured AGM reaggregates (4 independent experiments and 5 litters for uncultured AGMs: +9.5+/+ [n = 13]; +9.5+/− [n = 18]; +9.5−/− [n = 7]; 2 independent experiments and 2 litters for cultured AGM explants: +9.5+/+ [n = 5]; +9.5+/− [n = 13]; +9.5−/− [n = 3]; 5 independent experiments and 5 litters for cultured AGM reaggregates: +9.5+/+ [n = 10]; +9.5+/− [n = 20]; +9.5−/− [n = 12]). (E and G) Quantitative analysis of cell number in E11.5 uncultured AGMs (E) and cultured AGM explants (G). (F and H) Quantitative analysis of colony-forming activity of hematopoietic progenitors in E11.5 uncultured AGMs (F), cultured AGM explants (H), and cultured AGM reaggregates (H; 2 independent experiments and 2 litters for uncultured AGMs: +9.5+/+ [n = 5]; +9.5+/− [n = 10]; +9.5−/− [n = 4]; 2 independent experiments and 2 litters for cultured intact AGM explants: +9.5+/+ [n = 5]; +9.5+/− [n = 5]; +9.5−/− [n = 3]; 2 independent experiments and 2 litters for cultured AGM reaggregates: +9.5+/+ [n = 6]; +9.5+/− [n = 8]; +9.5−/− [n = 6]). All error bars represent SEM. *, P < 0.05; ***, P < 0.001 (two-tailed unpaired Student’s t test).](https://cdn.rupress.org/rup/content_public/journal/jem/210/13/10.1084_jem.20130733/4/jem_20130733r_fig1.jpeg?Expires=1767769257&Signature=5Jm~2w5fEuUTgghgKvJ8SvXZzHjNFoDpfaBca2HgmohPvXry857Rgt-klqkNumx1OkQ7etrXT4taCNau39xp35BY9~A3zhS~354hdlCdtzrCSYYlmALpZN89fL41r53dFLokTqrPfSd3qHSGcLnfMgfaORIQoMU4ZVmSpaem8TKnGGEHQHbRBopjEHPedWjsqhEdC1reL2xmr04aFBMveN5a9kHF8VVzAlMCQXya65cyfFhH1iuf4Yi7dLP4b5tki6G~3z1iYiv0kntd7AR~X5~VX74KNLG0Sax-qMpqt7VQUlXU1DpFmvF-GZjT4p7LDAUP76CaHrtNJ~9gqqlrkA__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
Cis-regulatory element requirement for Gata2 expression and hematopoietic progenitor activity in the AGM. (A) Schematic diagram for targeted deletion of the E-box-GATA composite element in murine Gata2 locus. (B) AGM ex vivo culture system. (C) Representative images of E11.5 +9.5+/+,+9.5+/−, and +9.5−/− embryos. (D) Real-time RT-PCR analysis of Gata2 expression levels in E11.5 uncultured AGMs, cultured AGM explants, and cultured AGM reaggregates (4 independent experiments and 5 litters for uncultured AGMs: +9.5+/+ [n = 13]; +9.5+/− [n = 18]; +9.5−/− [n = 7]; 2 independent experiments and 2 litters for cultured AGM explants: +9.5+/+ [n = 5]; +9.5+/− [n = 13]; +9.5−/− [n = 3]; 5 independent experiments and 5 litters for cultured AGM reaggregates: +9.5+/+ [n = 10]; +9.5+/− [n = 20]; +9.5−/− [n = 12]). (E and G) Quantitative analysis of cell number in E11.5 uncultured AGMs (E) and cultured AGM explants (G). (F and H) Quantitative analysis of colony-forming activity of hematopoietic progenitors in E11.5 uncultured AGMs (F), cultured AGM explants (H), and cultured AGM reaggregates (H; 2 independent experiments and 2 litters for uncultured AGMs: +9.5+/+ [n = 5]; +9.5+/− [n = 10]; +9.5−/− [n = 4]; 2 independent experiments and 2 litters for cultured intact AGM explants: +9.5+/+ [n = 5]; +9.5+/− [n = 5]; +9.5−/− [n = 3]; 2 independent experiments and 2 litters for cultured AGM reaggregates: +9.5+/+ [n = 6]; +9.5+/− [n = 8]; +9.5−/− [n = 6]). All error bars represent SEM. *, P < 0.05; ***, P < 0.001 (two-tailed unpaired Student’s t test).
