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. 2010 Apr 12;5(4):e10127. doi: 10.1371/journal.pone.0010127

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Hale et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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Bone marrow chimeras were generated by reconstituting irradiated wild-type Ly5.1⁺Ly5.2⁺ mice with bone marrow cells from either Lmna ^+/+ or Lmna ^-/- Ly5.2⁺Ly5.1^- donor mice. Single-cell suspensions from bone marrow, thymus, and spleen were analyzed 44 weeks following irradiation and bone marrow reconstitution. Cells were stained for surface Ly5.1, Ly5.2, B220, CD4, CD8, TCRβ, and CD19 expression. Donor-derived Ly5.2⁺Ly5.1^- cells were analyzed in thymus and spleen. Total bone marrow cells were analyzed because not all bone marrow cells express Ly5; however, >98% of Ly5⁺ cells were donor-derived Ly5.2⁺Ly5.1^-, indicating efficient engraftment. A. The percentage of total bone marrow cells that are B220⁺ (left) and the absolute number of total bone marrow cells and B220⁺ bone marrow cells (right) in the indicated bone marrow chimeric mice. B. The percent (left) and absolute number (right) of the indicated donor cells that are DN, DP, CD4 SP, and CD8 SP thymocytes. C. The percent (left) and absolute number (right) of the indicated donor cells that are CD4⁺ T cells, CD8⁺ T cells, and CD19⁺ B cells. Charts show the mean percent or number with error bars indicating the standard deviation (Lmna ^+/+ into wild-type (WT) chimera N = 3; Lmna ^-/- into WT chimera N = 5). P values were calculated using an unpaired two-tailed Student's t test and none reached significance.