American Society of Hematology

Figure 1

$Figure 1. Zbt16lu/lu HSCs present defects associated with aged HSCs. (A) Serial transplantation assay. Donor BM CD45.2 Zbtb16lu/lu or WT cells were injected into lethally irradiated CD45.1 recipient mice and were serially transplanted. Analyses were performed at 14 weeks or at 52 weeks after transplant. (B) Analysis of CD45.2+ donor cells in BM, spleen, thymus, and blood 14 weeks after each transplant. Representative fluorescence-activated cell sorting analyses (C) and quantitative and statistical analyses (D) of long-term reconstitution in LSK and stem cell compartments 14 weeks after secondary transplant. (E) Quantitative analysis of long-term reconstitution in progenitor cell compartment 14 weeks after tertiary transplant. (F) Quantitative analysis of the HSC compartment in 1-year-old Zbtb16lu/lu and WT mice. Bars represent mean ± SEM (n = 5). (G-H) Analysis of hematopoietic compartments 1 year after primary and secondary transplants. (G) Percentage of LT-HSC, ST-HSC, and MPP1 fractions among CD45.2+ HSCs 1 year after primary transplant. (H) Distribution of CD45.2+ myeloid and CD45.2+ B cells in BM and peripheral blood 1 year after secondary transplant. For 14-week post-reconstitution bar graphs, data represent mean ± SEM. Data are based on 2 (primary, secondary, and tertiary transplants) experimental repeats, with 6 recipient mice per group (n = 12). *P < .05; **P < .01; ***P < .001. For 52-week post-reconstitution bar graphs, data are represented as mean ± SEM (n = 6). *P < .05; **P < .01. (I) Competitive reconstitution analysis. Donor CD45.2 young Zbtb16lu/lu or WT aged BM cells were transplanted with an equal number of CD45.1/2 competitor WT BM cells into lethally irradiated CD45.1 recipients. Quantitative and statistical analyses of long-term reconstitution in stem cell compartments 21 weeks after transplant. Bars represent mean ± SEM (n = 6). CLP, common lymphoid progenitor; CMP, common myeloid progenitor; LT, long-term; MEP, megakaryocyte-erythroid progenitor cells; MPP, multipotent progenitor; SEM, standard error of the mean; ST, short-term; wks, weeks.$

Zbt16^lu/luHSCs present defects associated with aged HSCs. (A) Serial transplantation assay. Donor BM CD45.2 Zbtb16^lu/lu or WT cells were injected into lethally irradiated CD45.1 recipient mice and were serially transplanted. Analyses were performed at 14 weeks or at 52 weeks after transplant. (B) Analysis of CD45.2⁺ donor cells in BM, spleen, thymus, and blood 14 weeks after each transplant. Representative fluorescence-activated cell sorting analyses (C) and quantitative and statistical analyses (D) of long-term reconstitution in LSK and stem cell compartments 14 weeks after secondary transplant. (E) Quantitative analysis of long-term reconstitution in progenitor cell compartment 14 weeks after tertiary transplant. (F) Quantitative analysis of the HSC compartment in 1-year-old Zbtb16^lu/lu and WT mice. Bars represent mean ± SEM (n = 5). (G-H) Analysis of hematopoietic compartments 1 year after primary and secondary transplants. (G) Percentage of LT-HSC, ST-HSC, and MPP1 fractions among CD45.2⁺ HSCs 1 year after primary transplant. (H) Distribution of CD45.2⁺ myeloid and CD45.2⁺ B cells in BM and peripheral blood 1 year after secondary transplant. For 14-week post-reconstitution bar graphs, data represent mean ± SEM. Data are based on 2 (primary, secondary, and tertiary transplants) experimental repeats, with 6 recipient mice per group (n = 12). *P < .05; **P < .01; ***P < .001. For 52-week post-reconstitution bar graphs, data are represented as mean ± SEM (n = 6). *P < .05; **P < .01. (I) Competitive reconstitution analysis. Donor CD45.2 young Zbtb16^lu/lu or WT aged BM cells were transplanted with an equal number of CD45.1/2 competitor WT BM cells into lethally irradiated CD45.1 recipients. Quantitative and statistical analyses of long-term reconstitution in stem cell compartments 21 weeks after transplant. Bars represent mean ± SEM (n = 6). CLP, common lymphoid progenitor; CMP, common myeloid progenitor; LT, long-term; MEP, megakaryocyte-erythroid progenitor cells; MPP, multipotent progenitor; SEM, standard error of the mean; ST, short-term; wks, weeks.

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