Absence of HSCs from bone marrow of vav-cre NKAP cKO neonates. (A) Early hematopoiesis in total bone marrow single-cell suspensions from WT and vav-cre NKAP cKO neonates was analyzed for LSK (lin⁻Sca1⁺cKit⁺) and SLAM-HSC (lin⁻Sca1⁺cKit⁺CD48⁻CD150⁺). (B) Representative H&E staining of fixed bone marrow sections from WT and vav-cre NKAP cKO neonates is shown. Please note that at birth there is little accumulation of fat deposits, which develop with age. (C) Early hematopoietic progenitors were examined in liver from wild-type or NKAP cKO neonates. The resulting suspensions were stained with a lineage cocktail, c-Kit, Sca-1, Flt3, CD48, and CD150 to analyze LSK, as well as MPP (lin⁻Sca1⁺cKit⁺Flt3⁺), and SLAM-HSC (lin⁻Sca1⁺cKit⁺CD48⁻CD150⁺). (D) HSC (Lin⁻ckit^hiSca-1⁺CD27⁻), MPP (Lin⁻ckit^hiSca-1⁺CD27⁺⁾, CMP (Lin⁻ckit^hiSca-1⁻), CLP (Lin⁻IL7R⁺ckit^loSca-1⁺), and Pre-Pro B cells (CD43⁺B220⁺CD19⁻) were isolated by FACS, and expression of NKAP, GATA2 and 18S was analyzed by Q-PCR. Results shown are the average from triplicate amplifications of a single population sort, normalized to 18S, and then standardized between samples to the expression of either NKAP or GATA2 in the MPP population (= 1.0). Error bars represent SD.