Loss of HSCs and expansion of multipotent progenitors in +42M kb−/− mice. (A) SLAM CD48+CD150+ markers were used to characterize cell distribution within the MPP, LT-HSC, and ST-HSC cell populations gated from lin−Sca-1+c-kit+ (LSK) cell populations. (B) Absolute cell numbers for LSK, MPP, LT-HSCs, and ST-HSCs were calculated from bone marrow white cell count per femur. (C) Cebpa expression by RNA-seq (FPKM values of WT vs homozygous mice). (D) Total bone marrow cells from WT, heterozygous, and homozygous mice were cultured in semisolid medium supplemented with IL-3, IL-6, SCF, and GM-CSF. Colonies were counted and replated every 7 days. FACS plots showing that the majority of cells grown under these conditions are mainly LK/GMP cells and, to a lesser extent, MPP/LSK cells. Morphological examination with May-Grünwald-Giemsa after 7 days distinguishes normal granulocytic and macrophage differentiation in WT cells as compared with homozygous cells that show blasts as the major cell population.
