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. Author manuscript; available in PMC: 2021 Jul 1.
Published in final edited form as: Stem Cells. 2020 Mar 19;38(7):849–859. doi: 10.1002/stem.3174

Figure 1: Loss of CXCR4 in BM stromal cells reduces HSPC retention and function.

Figure 1:

(A) Top; Representative flow cytometry plots of freshly isolated mouse BM stromal cells (CD45-Ter119-), MSCs (CD45-Ter119-CD31-CD51+PDGFRα+) and ECs (CD45-Ter119-CD31+VE-cadherin+). Bottom; Representative flow plots showing CXCR4 expression on total stromal cells, MSC and EC. (B) Cell surface CXCR4 expression on mouse BM stromal cells, MSC, and EC (X±SEM; N= 5 mice, each assayed individually). (C) Schematic of chimeric mice generation, CXCR4 deletion and functional analysis. (D-E) CXCR4 transcript expression in CD45-Ter119-stromal cells of chimeric mice with WT stroma or CXCR4 KO stroma (X±SEM; N= 3 mice, each assayed individually) and cell surface CXCR4 expression on BM total stromal cells, MSC and EC of chimeric mice with WT stroma or CXCR4 KO stroma (X±SEM; N= 5 mice, each assayed individually). (F-G) LSK and SLAM-LSK frequency in the BM of chimeric mice with WT donor:CXCR4 KO stroma or WT donor:WT stroma at 12-week post tamoxifen treatment (X±SEM; N= 5 mice per group, assayed individually; *=P<0.05). (H) Cell-cycle status of BM SLAM-LSK from chimeric mice with WT or KO stroma (X±SEM; N= 4 mice per group, assayed individually; *=P<0.05). (I) Peripheral blood total CFU-C (CFU-GM, BFU-E, CFU-GEMM) in both group of mice at 12-week tamoxifen treatment (X±SEM; N= 4 mice per group assayed individually; *=P<0.05). (J) Long-term repopulating ability of HSPC from chimeric mice with WT donor:CXCR4 KO stroma or WT donor:WT stroma. BM cells from chimeric mice (CD45.1+) plus 200,000 competitive whole BM cells from untreated CD45.2+ mice were transplanted into lethally irradiated CD45.2+ recipients. Peripheral blood chimerism and multi-lineage reconstitution was assessed 20 weeks’ post-transplant (X±SEM; N= 4 mice per group, each assayed individually; *=P<0.05).