Heme exacerbates immune cell recruitment upon apoptotic damage through inflammatory macrophages in sickle cell disease. (A) Percentage of total phagocytes (i), resident and recruited phagocytes (ii,iii), Ly6C⁺ recruited monocytes (iv), and 7AAD⁺ dead macrophages (vi); representative flow cytometry plot of CD11b^low F4/80⁺ resident KCs and CD11b⁺ F4/80^+/− recruited phagocytes (v) in the liver of untreated HbA and HbS mice and HbA, HbS, and Hx-treated HbS mice receiving 1 × 10⁷ AC infusion. (B) Percentage of total phagocytes (i), resident and recruited phagocytes (ii,iii), and representative flow cytometry plot of phagocytes (iv) in the liver of untreated and heme-treated Wt mice receiving or not receiving 1 × 10⁷ AC infusion. (C) Number of F4/80^int/high monocytes/macrophages, Gr-1⁺ neutrophils and monocytes, F4/80⁻ Gr-1⁻ lymphocytes recruited to the peritoneum of Wt mice receiving BMDMs untreated (NT) or previously exposed for 15 hours to 5 μM heme bound to 5 μM albumin (Alb) or 5 μM Hx (i) and representative flow cytometry plot (ii). Data shown are average of 3 independent experiments. Values represent mean ± SEM. Statistical analysis was performed by comparing 3 or more groups with 1-way ANOVA followed by Bonferroni posttest. ∗P < .05; ∗∗P < .01; ∗∗∗P < .001; ∗∗∗∗P < .0001.