Figure 2.
Simultaneous lack of Jagged1 in the BM stroma and Notch1 in HSCs of mixed BM chimeras does not affect their self-renewal potential. (A) Mixed BM chimeric mice analyzed 14 to 15 weeks after reconstitution with a 1:2 mixture of B6-SJL (CD45.1+) and either J1lox/lox or J1-/- (CD45.2+) BM-derived populations. Percentage of CD45.2+ BM cells, CD45.2+Lin- cells, and HSCs (CD117+Sca1+) gated on lin-CD45.2+ cells from either J1lox/lox (▪) or J1-/- mice (▦). The bars represent mean ± SD (n = 8). (B) Mixed BM chimeric mice using either J1lox/lox or induced J1-/- recipients (both CD45.2+) analyzed 8 to 20 weeks after reconstitution with a 1:1 mixture of C57BL/6 (CD45.2+) and either N1lox/lox or N1-/- (CD45.1+) BM-derived populations. (Top row) Percentage of CD45.1+ BM cells, CD45.1+Lin- cells, and HSCs (CD117+Sca1+), long-term-HSCs (CD117+Sca1+CD135-), short-term-HSCs (CD117+Sca1+CD135+), CMPs (CD117+Sca1-), and CLPs (CD117intSca1int) gated on lin-CD45.1+ BM cells. (Bottom row) B cells (B220+), immature myeloid cells (CD11b+/Gr1lo/int), eosinophils (CD11b+/Gr1+), early erythroblasts (Ter119+) gated on CD45.1+ cells, and percentage of CD45.1+ cells in the thymus. Most of the thymic CD45.1+ cells derived from N1-/- BM cells are B cells. Control into control (▪; n = 8), control into J1-/- (▨; n = 3), N1-/- into control (▥; n = 7), or N1-/- into J1-/- (□; n = 14) mixed BM chimeras. The bars represent mean ± SD. (C) PCR analysis of the deletion of the N1 gene using genomic DNA from CD45.1+ HSCs sorted 8 weeks after transplantation of the indicated chimeric mice.