Hemolysis leads to IFN-I-dependent recruitment of monocytes to the liver. Frequencies of (A) CMo (gating strategy in supplemental Figure 2A-B) and (B) Ly-6C⁺MHC-II⁺ monocyte in total CD45⁺ leukocytes in blood, spleen, and liver of control mice and sickle mice (n = 3-4). Frequencies of (C) CMo and (D) Ly-6C⁺MHC-II⁺ monocyte in total CD45⁺ leukocytes in blood and liver of WT mice and ifnar1^−/− mice at 20 hours after IV injection with PBS as control (200 µL/20 g body weight) or hemin (35 µmol/kg body weight) (n = 4-5). (E) Plasma CCL2 levels in control mice and sickle mice (n = 4-5). (F) Plasma CCL2 levels in mice shown in panel C (n = 4-5). (G) Representative dot plot and histogram showing the gating strategy for analysis of total Mⲫ (F4/80⁺CD11b^low Mⲫ), monocyte derived Mⲫ (Tim-4⁻ MoMⲫ), and resident Mⲫ (Tim-4⁺ Mⲫ) in mouse liver and spleen. (H) Frequencies of MoMⲫ in total Mⲫ of spleen and liver in control and sickle mice (n = 3-6). (I) Representative histograms showing the gating strategy for liver Tim-4⁻ MoMⲫ, and Tim-4⁺ Mⲫ in WT mice, ifnar1^−/− mice, and ccr2^−/− mice at 72 hours after IV injection with PBS as control (200 µL/20 g body weight) or hemin (35 µmol/kg body weight). (J) Frequencies of MoMⲫ in total Mⲫ in livers of mice as shown in I (n = 3-6). Means were compared using 2-tailed Student t test. *P < .05.