Fig. 4

NOD1 depletion corrects diabetes-induced hematopoietic imbalance. A Flow cytometry gating strategy for bone marrow single-cell suspensions. Viable/live singlets were gated on Lin⁻CD127^± cells. Representative FACS plots of the analysis of HSPC subpopulations gated as LSK (Lin⁻CD127⁻/Sca1⁺c-Kit⁺), LT-HSC (Lin⁻CD127⁻/Sca1⁺c-Kit⁺/CD34⁻CD135⁻), ST-HSC (Lin⁻CD127⁻/Sca1⁺c-Kit⁺/CD34⁺CD135⁻), CLP (Lin⁻CD127⁻/Sca1^medc-Kit^med), CMP (Lin⁻CD127⁻Sca1⁻c-Kit⁺CD34⁺FcgRII/III^lo), MEP (Lin⁻CD127⁻Sca1⁻c-Kit⁺CD34⁻FcgRII/III^lo) and GMP (Lin⁻CD127⁻Sca1⁻c-Kit⁺CD34⁺FcgRII/III^hi) (n = 11–16 per group). B Representative flow cytometry diagrams of LT- and ST-HSCs in each experimental group. C–E Percentages of bone marrow LSK, LT-HSCs and ST-HSCs out of total live cells. F The ratio of the percentages of ST-HSCs to LT-HSCs in bone marrow. G, H Representative flow cytometry diagrams of CMP and CLP cells in each experimental group. I–J Percentages of bone marrow CMP and CLP cells out of total live cells. K CFU assay results, i.e., quantification of the numbers of CFU-G/M/GMs after plating bone marrow cells. (n = 15 per group). The results are presented as the mean ± SD; *p < 0.05, **p < 0.01. WT, wild type; HSPC, hematopoietic stem/progenitor cell; LT-HSC, long-term repopulating hematopoietic stem cell; ST-HSC, short-term repopulating hematopoietic stem cell; CLP, common lymphoid progenitor; CMP, common myeloid progenitor; GMP, granulocyte–macrophage progenitor; MEP, megakaryocyte erythroid progenitor