ARAP3 R302,303A mutation impairs HSC function.

<p>(A) Complete blood counts (CBC) of <i>f/f</i> control mice (white bars) and <i>KI/KI</i> mutant mice (dotted black bars). n = 4. (B) Colony-forming cell (CFC) assays of <i>f/f</i> and <i>KI/KI</i> bone marrow cells enumerated after 11 days in culture. Colonies identified as multipotent myeloid progenitors (GEMM), erythroid (BFU-E), granulocyte-monocyte progenitors (GM), granulocyte (G), or monocyte (M). n = 3. (C) The percentage of SLAM LSK population enriched for long-term hematopoietic stem cells in <i>f/f</i> and <i>KI/KI</i> bone marrow, gated on the Lin⁻c-Kit⁺Sca1⁺ (LSK) population, followed by SLAM markers CD48⁻CD150⁺. (D–K) Transplantation of LSK cells from <i>f/f</i> and <i>KI/KI</i> donor mice. Results were pooled from two separate experiments. (D) Peripheral blood in the recipients was assessed at 4, 8, and 12 weeks after primary transplant by flow cytometry for the percentage of donor-derived leukocytes. (E,H) Donor contribution to myeloid, B-cell, and T-cell compartments in the peripheral blood at the end of the primary (E) or secondary (H) transplant was analyzed by flow cytometry. (F,H) Donor percentages in the total BM, LSK, and SLAM LSK compartments (CD48⁻CD150⁺LSK) were enumerated by flow cytometry, 16 weeks post primary (F) or secondary (H) transplantation. (I) Peripheral blood reconstitution in serial transplants was assayed by flow cytometry. 1°: primary; 2°: secondary; 3°: tertiary BMT. (J,K) Bars show lineage distributions within donor-derived cells of individual recipient mice (left Y-axis), while diamond symbols indicate total donor leukocyte percentages (right Y-axis) in the peripheral blood at the end of the primary (J) or secondary (K) BM transplants. T: CD3⁺; B: CD19⁺; M: Mac1⁺. Graphs show mean ±SEM. P-values determined by two-tailed Student's t-test.