Figure 2.
Canonical Notch signaling is dispensable for stem and myelo-E progenitor cell replenishment in competitive bone marrow chimeras. (A-C) One million BM cells were harvested from 10- to 14-week-old Rbpjfl/flMx1Cre/+ and control (Rbpjfl/flMx1+/+ and Rbpjfl/+Mx1Cre/+) mice (CD45.2) 4 to 5 weeks after poly(I:C) treatment and transplanted into lethally irradiated wild-type (WT; CD45.1 or CD45.1/2) recipients together with 1 million competitor WT (CD45.1 or CD45.1/2) adult BM cells. Reconstitution of HSC (N = 2 of Rbpjfl/flMx1Cre/+ and N = 1 and N = 3 of Rbpjfl/flMx1+/+ and Rbpjfl/+Mx1Cre/+, respectively) and myeloid progenitor subsets (N = 5 of Rbpjfl/flMx1Cre/+, N = 2 of Rbpjfl/flMx1+/+, N = 3 of Rbpjfl/+Mx1Cre/+) in the BM of engrafted mice was assessed 7 to 9 weeks after transplantation. Percentage donor (CD45.2)-derived reconstitution of (A) HSC (mean ± SD) and (B) myeloid progenitor subsets (mean ±SEM) relative to total BM cells. (C) Mean (SEM) CD45.2 contribution of Rbpjfl/flMx1Cre/+ (N = 6) and control (N = 2 and N = 6 of Rbpjfl/flMx1+/+ and Rbpjfl/+Mx1Cre/+, respectively) BM cells to total NK1.1−Mac1+ myeloid, NK1.1−Mac1−CD19+ B and NK1.1−Mac1−CD4/CD8+ T cells. (D) Reconstitution of CD45.2-derived blood platelets (CD41+CD150+eGFP−) of total platelets in Vwf-eGFP (CD45.1/2) recipients 4 to 5 weeks after transplantation with 1 million 10- to 11-week-old CD45.2 Rbpjfl/flMx1Cre/+ (N = 4) or control Rbpjfl/+Mx1Cre/+ (N = 6) BM cells and 1 million Vwf-eGFP (CD45.1/2) competitor BM cells. (Left) Representative FACS profiles of platelet reconstitution in engrafted mice. (Right) Mean (SEM) percentage of CD150+CD41+ platelets derived from transplanted CD45.2 BM cells. For all data sets (A-D), statistical significance was investigated between Rbpj-deleted and control mice. ***P < .001.