Figure 2.
Defective hematopoietic reconstitution in mice that received transplantation with NFAT5-deficient bone marrow or HSCs. (A) Schematic diagram of the experiment (upper left), and donor-derived (CD45.2+) reconstitution of blood, different bone marrow populations, and hematopoietic stem and progenitor subsets in mice that received transplantation with bone marrow (0.5 × 106 cells) from hematopoietic cell–specific NFAT5-deficient mice (Nfat5fl/fl Vav-Cre) or littermate WT mice. Results are from 3 independent experiments, each with 1 bone marrow donor of the respective genotype and 6 to 9 recipients. (B) Donor-derived reconstitution of bone marrow hematopoietic stem and progenitor subsets in mice that received transplantation with bone marrow from myeloid-specific NFAT5-deficient mice (Nfat5fl/fl LysM-Cre) or littermate WT mice (Nfat5+/+ LysM-Cre). Results are from 2 independent experiments (left panel) or 1 experiment (right panel), each with 1 bone marrow donor of each genotype and 3 to 7 recipient mice. (C) Schematic diagram of transplants with Nfat5fl/fl Vav-Cre or WT HSCs together with competitor WT bone marrow cells. For primary transplants, fluorescence-activated cell sorter (FACS)–sorted CD45.2+Nfat5fl/fl Vav-Cre or WT HSCs were mixed with competitor WT CD45.1.2+ bone marrow cells and transplanted into irradiated CD45.1+ recipient mice. For secondary and tertiary transplants, donor cells were from whole bone marrow. (D) Competitor (CD45.1.2+) vs donor (CD45.2+) chimerism in peripheral blood cells isolated from mice serially reconstituted from NFAT5-deficient and control HSCs in competition with WT bone marrow. Blood samples were obtained 4 and 12 weeks after transplant. See supplemental Figure 5A-B for the number and percentage of donor- and competitor-derived blood cells. (E) Numbers of CD45.2+ donor- and (F) CD45.1.2+ competitor- derived reconstitution of bone marrow hematopoietic stem and progenitor subsets through serial transplants derived from Nfat5fl/fl Vav-Cre or WT HSCs. Analysis was done at week 13 after each transplant. (G) Competitor (CD45.1.2+) vs donor (CD45.2+) chimerism in hematopoietic stem and progenitor subsets isolated from mice serially reconstituted from NFAT5-deficient and control HSCs in competition with WT bone marrow. Results in panels D-G are from 2 independent transplant experiments, each with 1 HSC donor of each genotype and 3 to 6 recipients for the primary transplants. Secondary and tertiary transplants were done with pools of reconstituted bone marrow (4-6 WT and 3-5 NFAT5-deficient donors were pooled for the secondary transplant; and 5-7 WT and 3-8 NFAT5-deficient donors were pooled for the tertiary transplant). Results are shown as mean ± SEM. Statistical test: unpaired t test. ∗P < .05, ∗∗P < .01, ∗∗∗P < .001. P values < .1 are also indicated.

Defective hematopoietic reconstitution in mice that received transplantation with NFAT5-deficient bone marrow or HSCs. (A) Schematic diagram of the experiment (upper left), and donor-derived (CD45.2+) reconstitution of blood, different bone marrow populations, and hematopoietic stem and progenitor subsets in mice that received transplantation with bone marrow (0.5 × 106 cells) from hematopoietic cell–specific NFAT5-deficient mice (Nfat5fl/fl Vav-Cre) or littermate WT mice. Results are from 3 independent experiments, each with 1 bone marrow donor of the respective genotype and 6 to 9 recipients. (B) Donor-derived reconstitution of bone marrow hematopoietic stem and progenitor subsets in mice that received transplantation with bone marrow from myeloid-specific NFAT5-deficient mice (Nfat5fl/fl LysM-Cre) or littermate WT mice (Nfat5+/+ LysM-Cre). Results are from 2 independent experiments (left panel) or 1 experiment (right panel), each with 1 bone marrow donor of each genotype and 3 to 7 recipient mice. (C) Schematic diagram of transplants with Nfat5fl/fl Vav-Cre or WT HSCs together with competitor WT bone marrow cells. For primary transplants, fluorescence-activated cell sorter (FACS)–sorted CD45.2+Nfat5fl/fl Vav-Cre or WT HSCs were mixed with competitor WT CD45.1.2+ bone marrow cells and transplanted into irradiated CD45.1+ recipient mice. For secondary and tertiary transplants, donor cells were from whole bone marrow. (D) Competitor (CD45.1.2+) vs donor (CD45.2+) chimerism in peripheral blood cells isolated from mice serially reconstituted from NFAT5-deficient and control HSCs in competition with WT bone marrow. Blood samples were obtained 4 and 12 weeks after transplant. See supplemental Figure 5A-B for the number and percentage of donor- and competitor-derived blood cells. (E) Numbers of CD45.2+ donor- and (F) CD45.1.2+ competitor- derived reconstitution of bone marrow hematopoietic stem and progenitor subsets through serial transplants derived from Nfat5fl/fl Vav-Cre or WT HSCs. Analysis was done at week 13 after each transplant. (G) Competitor (CD45.1.2+) vs donor (CD45.2+) chimerism in hematopoietic stem and progenitor subsets isolated from mice serially reconstituted from NFAT5-deficient and control HSCs in competition with WT bone marrow. Results in panels D-G are from 2 independent transplant experiments, each with 1 HSC donor of each genotype and 3 to 6 recipients for the primary transplants. Secondary and tertiary transplants were done with pools of reconstituted bone marrow (4-6 WT and 3-5 NFAT5-deficient donors were pooled for the secondary transplant; and 5-7 WT and 3-8 NFAT5-deficient donors were pooled for the tertiary transplant). Results are shown as mean ± SEM. Statistical test: unpaired t test. ∗P < .05, ∗∗P < .01, ∗∗∗P < .001. P values < .1 are also indicated.

Close Modal
This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal