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. 2018 Mar 15;7(6):468–476. doi: 10.1002/sctm.17-0189

Figure 4.

Figure 4

“11a/EPCR” two‐color sorting is as efficient as using the “SLAM” method. (A): Representation of direct comparison of two‐color sorting methods. BM from CFP or Tomato mice was sorted using the combination of CD11a and EPCR only or the combination of CD150 and CD48 only. Approximately 380 cells from each of CD11a– EPCR+ and CD150+ CD48– gates were sorted, mixed, and co‐transplanted with added 250,000 helper/competitor WBM (Group 1 recipients). Percentages of cells within each gate was kept consistent between the two methods. 200,000 cells from outside of the CD11a– EPCR+ gate (“Not 11a/EPCR”) and outside of the CD150+ CD48– gate (“Not SLAM”) were also mixed and co‐transplanted (Group 2 recipients). In (B, C), CFP+ donor‐derived cells are represented by outlined symbols and Tomato+ donor‐derived cells with borderless symbols. (Bi): Time‐course analysis of blood granulocyte chimerism from CD11a– EPCR+ and CD150+ CD48– sources in primary recipients 4, 9, and 12 weeks (W) post‐transplant, and in secondary recipients (separated by vertical dashed line) at week 6 following secondary transplant. Primary recipients used for secondary transplants are marked with an “x” inside circles at the 12‐week timepoint. (Bii): Donor chimerism of HSPCs in the BM of primary recipients transplanted with CD11a– EPCR+ and CD150+ CD48– sorted cells 13 weeks post‐transplant. (Ci): Time‐course analysis of blood granulocyte chimerism from “Not 11a/EPCR” and “Not SLAM” sources in primary recipients 4, 9, and 12 weeks (W) post‐transplant, and in secondary recipients (separated by vertical dashed line) at week 6 following secondary transplant. The primary recipients used for secondary transplant is marked (half shaded black) at the 12‐week timepoint. (Cii): Donor chimerism of HSPCs in the BM of primary recipients transplanted with “Not 11a/EPCR” and “Not SLAM” sorted cells 13 weeks post‐transplant. Number of experiments = 2. **, p ≤ .01; ***, p ≤ .001 (Student's unpaired t test). “Not 11a/EPCR” = not CD11a‐EPCR+; “Not SLAM” = not CD150+CD48–. Abbreviations: BM, bone marrow, EPCR, endothelial protein C receptor; HSPC, hematopoietic stem and progenitor cells.