Defective SEP differentiation in Irg1-deficient mice delayed the recovery from HKBA-induced inflammatory anemia. (A-G) Age- and sex-matched WT and Irg1–/– mice were administered with HKBA (5 × 108 particles per mouse) via intraperitoneal injection. (A) In the following 28 days, mice were monitored daily for survival and health, and blood was collected retro-orbitally every other day for hematocrit measurement (WT, n = 12; Irg1–/–, n = 8; repeated measures 2-way ANOVA followed by unpaired t test). (B-C) Analysis of Hb (left) and RBC counts (right) concentrations (B), and measurement of spleen weight (left) and splenocyte numbers (right) (C) at indicated time points after HKBA injection (n = 4 per group; unpaired t test). (D) A representative flow cytometry plot shows the gating of SEPs in the spleen isolated at day 8 after HKBA injection. (E-G) Analysis of percentages of Kit+Sca1–CD34–CD133– SEPs (E), frequency (top) and total numbers (bottom) of stress BFU-E colony formation (F), and Nos2 mRNA abundance (G) at indicated time points (n = 4 per group; unpaired t test). Data represent mean ± SEM. ∗P < .05; ∗∗P < .01; ∗∗∗P < .001. Hb, hemoglobin; RBC, red blood cell; Q1, quadrant 1.
