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. 2012 Mar 12;209(3):537–549. doi: 10.1084/jem.20110994

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© 2012 Mansour et al.

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Figure 5. — Restoration of OCL activity rescues LSK homing in the BM of oc/oc mice. Sorted CD45⁺Sca1⁺ hematopoietic progenitors from actin-GFP (3–4-wk-old) mice were transferred into newborn oc/oc and wild-type mice. Analyses were performed at day 45 for the wild-type and treated oc/oc mice (oc/oc + CD45⁺Sca1⁺), and at day 17 for the untreated oc/oc mice because of their short life span. (A) Typical FACS profile of sorted GFP⁺CD45⁺Sca1⁺ cells before transfer into irradiated recipient mice. (B) Radiological analysis of the femur of treated and control mice, representative of four mice per group in two independent experiments. (C) Flow cytometry analysis of CD45 and GFP expression in the BM of the wild-type and treated oc/oc mice. Percentage of the cells in each quadrant is indicated. Data are representative four mice in each group in two independent experiments. (D) Flow cytometry analysis of LSK cells in the BM of the treated +/+ and oc/oc mice and control untreated oc/oc mice. Cells were gated on Lin^neg cells. Percentages of the populations are indicated. Data are representative four mice per group in two independent experiments. (E) Bar graph shows the mean ± SD of the percentage of LSK cells in the BM and the spleen obtained for four mice per group in two independent experiments. *, P < 0.01 (+/+ versus control oc/oc mice); **, P < 0.01 (treated oc/oc versus control oc/oc mice). (F) Mean ± SD of the percentage of Sca1⁺c-kit^neg cells in Lin^neg cells from the BM and the spleen of four mice per group in two independent experiments. *, P < 0.01. (G) Flow cytometry analysis on BM LSK cells of 17-d-old PBS- or DC-treated oc/oc mice gated on lin^neg cells (top) and on CD45 expression gated on BM Lin^negSca1⁺c-kit^neg cells (bottom). Data are representative of those obtained for three mice per group in one experiment. (H) Flow cytometry analysis of BM Ter119^negCD45^neg mesenchymal cells. Data are representative of those obtained for three mice per group in one experiment. (I) Analysis of GFP expression in LSK cells from the wild-type and oc/oc mice treated with CD45⁺Sca1⁺GFP⁺ cells. Cells were gated on BM LSK cells. Bar graph shows the mean ± SD of the percentage of GFP⁺LSK cells obtained for four mice per group in two independent experiments. *, P < 0.05. (J) Flow cytometry analysis of the main hematopoietic lineages (B220 for B cells, CD11b for monocytes, and CD3 for T cells) in the BM of oc/oc mice treated with CD45⁺Sca1⁺GFP⁺ cells. Percentage of cells in each quadrant is indicated. Data are representative of four treated oc/oc mice in two independent experiments.