![Neutralization of IFNAR attenuates the reconstitution defects of NFAT5-deficient hematopoietic progenitors upon γ irradiation or treatment with 5-FU. (A) Percentage of the indicated hematopoietic progenitor populations in the bone marrow of mice of the different genotypes 55 days after TBI (5 Gy). WT (Nfat5+/+ Vav-Cre, Ifnar1+/+), hematopoietic cell–specific NFAT5-deficient (Nfat5fl/fl Vav-Cre, Ifnar1+/+), IFNAR1-deficient (Nfat5+/+ Vav-Cre, Ifnar1–/–), and double deficient (Nfat5fl/fl Vav-Cre, Ifnar1–/–) mice. Results are shown as mean ± SEM (n = 6-12 mice of each genotype). (B) Percentage of hematopoietic progenitor populations in the bone marrow of mice of the indicated genotypes treated with 3 consecutive doses of 5-FU (150 mg/kg) administered every 14 days, and analyzed 14 days after the last dose. Results are shown as mean ± SEM (n = 4-7 mice of each genotype). (C) Effect of neutralizing IFNAR1 during a 3-dose 5-FU regime on the percentage of hematopoietic progenitor populations. Mice were treated with control IgG or neutralizing anti-IFNAR1 antibody as described in “Materials and methods,” and analyzed 14 days after the last dose of 5-FU. Results are shown as mean ± SEM (n = 5 mice per condition). (D) Percentage of apoptotic cells (annexin V–positive [annexin V+] DAPI− [4′,6-diamidino-2-phenylindole–negative]) in hematopoietic progenitor populations in the bone marrow of mice of the indicated genotypes 55 days after TBI (5 Gy). Results are shown as mean ± SEM (n = 5-9 mice of each genotype). Statistical test: unpaired t test. ∗P < .05, ∗∗P < .01, ∗∗∗P < .001. P values < .1 are also indicated.](https://ash.silverchair-cdn.com/ash/content_public/journal/bloodadvances/8/21/10.1182_bloodadvances.2023011306/1/blooda_adv-2023-011306-gr5.jpeg?Expires=1733469344&Signature=POjbANJFxuxOIPMQBEgb0pEKtx-vnMfGvMjWRWdaE0BqQl8FibPoeI9eJK2yRL4nDYB14rORZofqRjUUkzBZ2B3oJut6Ps-glVLErAl0whKCwmph7Xl-8AsQBX9haPz2ttkilUfl-A2OU3hupYTsWuXpa8nWdLxTrgCvcmJ4oNfw~ftlQW9ZbcvT195k8UjWbz3~U9rz0~YUYjeQmeS~4SOAi1SS-4YgGYZNmIZFkoCty-1iJXoJojwfBxEKhG9xPTxYbEN-l8A4zcW0arJZJyD-RMCB828zB0QGbQkLrXX2pro1SkGrRungfeyT3K6ctloi2D~aYfRs44jVmL6Vsg__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
Neutralization of IFNAR attenuates the reconstitution defects of NFAT5-deficient hematopoietic progenitors upon γ irradiation or treatment with 5-FU. (A) Percentage of the indicated hematopoietic progenitor populations in the bone marrow of mice of the different genotypes 55 days after TBI (5 Gy). WT (Nfat5+/+ Vav-Cre, Ifnar1+/+), hematopoietic cell–specific NFAT5-deficient (Nfat5fl/fl Vav-Cre, Ifnar1+/+), IFNAR1-deficient (Nfat5+/+ Vav-Cre, Ifnar1–/–), and double deficient (Nfat5fl/fl Vav-Cre, Ifnar1–/–) mice. Results are shown as mean ± SEM (n = 6-12 mice of each genotype). (B) Percentage of hematopoietic progenitor populations in the bone marrow of mice of the indicated genotypes treated with 3 consecutive doses of 5-FU (150 mg/kg) administered every 14 days, and analyzed 14 days after the last dose. Results are shown as mean ± SEM (n = 4-7 mice of each genotype). (C) Effect of neutralizing IFNAR1 during a 3-dose 5-FU regime on the percentage of hematopoietic progenitor populations. Mice were treated with control IgG or neutralizing anti-IFNAR1 antibody as described in “Materials and methods,” and analyzed 14 days after the last dose of 5-FU. Results are shown as mean ± SEM (n = 5 mice per condition). (D) Percentage of apoptotic cells (annexin V–positive [annexin V+] DAPI− [4′,6-diamidino-2-phenylindole–negative]) in hematopoietic progenitor populations in the bone marrow of mice of the indicated genotypes 55 days after TBI (5 Gy). Results are shown as mean ± SEM (n = 5-9 mice of each genotype). Statistical test: unpaired t test. ∗P < .05, ∗∗P < .01, ∗∗∗P < .001. P values < .1 are also indicated.
