American Society of Hematology

Figure 5.

$TIMPless HSCs have reduced B-lymphopoiesis potential in vivo. (A) Schematic showing the experimental procedure for WT or QT BM transplant into WT mice. WT recipient were irradiated twice and 2 million BM cells from donor mice were injected IV. Recipient BM were analyzed by flow cytometry 16 weeks later. (B) Total BM cells (left panel) and donor-derived (CD45.2+) total BM cell (right panel), 16 weeks posttransplantation, in recipient mice (WT→WT, n = 5; QT→WT, n = 4). Data represent mean plus or minus SEM; groups were compared by the unpaired Student t test; *P < .05, **P < .01, and ***P < .001. (C) Percentage (left panel) and absolute count (right panel) of donor-derived (CD45.2+), CD11b+Gr-1+ myeloid cells, B cells, pre-B and pro-B cells, from the experiment in panel B. Data represent mean plus or minus SEM; groups were compared by the unpaired Student t test; *P < .05, **P < .01, and ***P < .001. (D) Percentage (left) and absolute number (right) of cells in the B-cell developmental fractions multilineage progenitor (MLP), CLP , Fr-A, Fr-B and C, Fr-C′ of transplanted cells (CD45.2+), from the experiment in panel B. Data represent mean plus or minus SEM; groups were compared by 2-way ANOVA; *P < .05. (E) Schematic showing the experimental procedure for secondary BM transplant. Transplanted BM of WT or QT from WT primary recipient subsequently injected into irradiated secondary WT recipient. Total 2 million BM cells transplanted into secondary recipient and analyzed by flow cytometry 16 weeks later. (F) Total BM cells (left panel) and donor-derived (CD45.2+) total BM cells count (right panel), 16 weeks postsecondary transplantation, in recipient mice (WT→WT→WT, n = 4; QT→WT→WT, n = 5). Data represent mean plus or minus SEM; WT or QT groups were compared by the unpaired Student t test; *P < .05. (G) Percentage (left panel) and absolute count (right panel) of donor-derived (CD45.2+), CD11b+Gr-1+ myeloid cells, B cells, pre-B-cell and pro-B cell, from the experiment in panel F. Data represent mean plus or minus SEM; groups were compared by the unpaired Student t test; *P < .05. (H) Percentage (left) and absolute number (right) of B-cell developmental fractions MLP, CLP, Fr-A, Fr-B and C, Fr-C′ of transplanted cells in recipient mice from the experiment in panel F. Data represent mean plus or minus SEM; WT and QT groups were compared by 2-way ANOVA.$

TIMPless HSCs have reduced B-lymphopoiesis potential in vivo. (A) Schematic showing the experimental procedure for WT or QT BM transplant into WT mice. WT recipient were irradiated twice and 2 million BM cells from donor mice were injected IV. Recipient BM were analyzed by flow cytometry 16 weeks later. (B) Total BM cells (left panel) and donor-derived (CD45.2⁺) total BM cell (right panel), 16 weeks posttransplantation, in recipient mice (WT→WT, n = 5; QT→WT, n = 4). Data represent mean plus or minus SEM; groups were compared by the unpaired Student t test; *P < .05, **P < .01, and ***P < .001. (C) Percentage (left panel) and absolute count (right panel) of donor-derived (CD45.2⁺), CD11b⁺Gr-1⁺ myeloid cells, B cells, pre-B and pro-B cells, from the experiment in panel B. Data represent mean plus or minus SEM; groups were compared by the unpaired Student t test; *P < .05, **P < .01, and ***P < .001. (D) Percentage (left) and absolute number (right) of cells in the B-cell developmental fractions multilineage progenitor (MLP), CLP , Fr-A, Fr-B and C, Fr-C′ of transplanted cells (CD45.2⁺), from the experiment in panel B. Data represent mean plus or minus SEM; groups were compared by 2-way ANOVA; *P < .05. (E) Schematic showing the experimental procedure for secondary BM transplant. Transplanted BM of WT or QT from WT primary recipient subsequently injected into irradiated secondary WT recipient. Total 2 million BM cells transplanted into secondary recipient and analyzed by flow cytometry 16 weeks later. (F) Total BM cells (left panel) and donor-derived (CD45.2⁺) total BM cells count (right panel), 16 weeks postsecondary transplantation, in recipient mice (WT→WT→WT, n = 4; QT→WT→WT, n = 5). Data represent mean plus or minus SEM; WT or QT groups were compared by the unpaired Student t test; *P < .05. (G) Percentage (left panel) and absolute count (right panel) of donor-derived (CD45.2⁺), CD11b⁺Gr-1⁺ myeloid cells, B cells, pre-B-cell and pro-B cell, from the experiment in panel F. Data represent mean plus or minus SEM; groups were compared by the unpaired Student t test; *P < .05. (H) Percentage (left) and absolute number (right) of B-cell developmental fractions MLP, CLP, Fr-A, Fr-B and C, Fr-C′ of transplanted cells in recipient mice from the experiment in panel F. Data represent mean plus or minus SEM; WT and QT groups were compared by 2-way ANOVA.

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