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. 2008 Dec 12;113(13):2924–2933. doi: 10.1182/blood-2008-02-138008

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© 2009 by The American Society of Hematology

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In situ deletion of SHIP does not compromise the capacity of HSCs to mediate long-term multilineage repopulation. (A) Contour plots for CD45.1 versus CD45.2 staining illustrating equal repopulation in PB at 60 days after transplantation before poly I:C treatment in 2 representative MxCreSHIP^flox/flox (CD45.2):WT-Ly5.1 (CD45.1) BM chimeras. (B) Percentage of donor repopulation in the PB by MxCreSHIP^flox/flox (CD45.2) and WT-Ly5.1 (CD45.1) HSCs in MxCreSHIP^flox/flox (CD45.2):WT-Ly5.1 (CD45.1) BM chimeras at 60 days after transplantation before poly I:C treatment. (C) Representative SHIP Western blot from single-positive CD45.1⁺CD45.2⁻- or CD45.2⁺CD45.1⁻-sorted cells isolated from splenocytes from MxCreSHIP^flox/flox (CD45.2):WT-Ly5.1 (CD45.1) BM chimeras 5 months after poly I:C treatment. Single-positive splenocytes were sorted and lysates were probed for SHIP expression. (D) Percentage of global repopulation in the PB by MxCreSHIP^flox/flox (CD45.2) and WT-Ly5.1 (CD45.1) HSC in MxCreSHIP^flox/flox (CD45.2):WT-Ly5.1 (CD45.1) BM chimeras at the indicated times after poly I:C treatment (n ≥ 5). At monthly intervals, the level of donor reconstitution was assessed in PB. (E) RU activity by MxCreSHIP^flox/flox (CD45.2) and WT-Ly5.1 (CD45.1) HSCs in the MxCreSHIP^flox/flox (CD45.2):WT-Ly5.1 (CD45.1) BM chimeras 5 months after poly I:C treatment (n ≥ 5). (F) Contour plots for CD45.1 versus CD45.2 staining that illustrate multilineage repopulation in PB 20 weeks after poly I:C treatment in a representative MxCreSHIP^flox/flox (CD45.2):WT-Ly5.1 (CD45.1) BM chimera. (G) Percentage repopulation of the indicated lymphoid and myeloid cell lineages by MxCreSHIP^flox/flox (CD45.2) and WT-Ly5.1 (CD45.1) HSC in MxCreSHIP^flox/flox (CD45.2):WT-Ly5.1 (CD45.1) BM chimeras 5 months after poly I:C treatment (n ≥ 5). At death, the level of donor reconstitution was assessed in BM. Significance was established using the unpaired Student t test (* P < .05). Errors shown represent the SEM; ■, cells derived from MxCreSHIP^flox/flox BM; , cells derived from WT-Ly5.1 BM.

Inline graphic — In situ deletion of SHIP does not compromise the capacity of HSCs to mediate long-term multilineage repopulation. (A) Contour plots for CD45.1 versus CD45.2 staining illustrating equal repopulation in PB at 60 days after transplantation before poly I:C treatment in 2 representative MxCreSHIP^flox/flox (CD45.2):WT-Ly5.1 (CD45.1) BM chimeras. (B) Percentage of donor repopulation in the PB by MxCreSHIP^flox/flox (CD45.2) and WT-Ly5.1 (CD45.1) HSCs in MxCreSHIP^flox/flox (CD45.2):WT-Ly5.1 (CD45.1) BM chimeras at 60 days after transplantation before poly I:C treatment. (C) Representative SHIP Western blot from single-positive CD45.1⁺CD45.2⁻- or CD45.2⁺CD45.1⁻-sorted cells isolated from splenocytes from MxCreSHIP^flox/flox (CD45.2):WT-Ly5.1 (CD45.1) BM chimeras 5 months after poly I:C treatment. Single-positive splenocytes were sorted and lysates were probed for SHIP expression. (D) Percentage of global repopulation in the PB by MxCreSHIP^flox/flox (CD45.2) and WT-Ly5.1 (CD45.1) HSC in MxCreSHIP^flox/flox (CD45.2):WT-Ly5.1 (CD45.1) BM chimeras at the indicated times after poly I:C treatment (n ≥ 5). At monthly intervals, the level of donor reconstitution was assessed in PB. (E) RU activity by MxCreSHIP^flox/flox (CD45.2) and WT-Ly5.1 (CD45.1) HSCs in the MxCreSHIP^flox/flox (CD45.2):WT-Ly5.1 (CD45.1) BM chimeras 5 months after poly I:C treatment (n ≥ 5). (F) Contour plots for CD45.1 versus CD45.2 staining that illustrate multilineage repopulation in PB 20 weeks after poly I:C treatment in a representative MxCreSHIP^flox/flox (CD45.2):WT-Ly5.1 (CD45.1) BM chimera. (G) Percentage repopulation of the indicated lymphoid and myeloid cell lineages by MxCreSHIP^flox/flox (CD45.2) and WT-Ly5.1 (CD45.1) HSC in MxCreSHIP^flox/flox (CD45.2):WT-Ly5.1 (CD45.1) BM chimeras 5 months after poly I:C treatment (n ≥ 5). At death, the level of donor reconstitution was assessed in BM. Significance was established using the unpaired Student t test (* P < .05). Errors shown represent the SEM; ■, cells derived from MxCreSHIP^flox/flox BM; , cells derived from WT-Ly5.1 BM.