Figure 6.

Delayed clearance of apoptotic neutrophils in the absence of Ly6C^hi monocytes and monocyte-derived macrophages (MoMF). (A) RNA-seq heat map analysis displaying the differential expression (z-scored) of genes between “MC-21” and “phosphate-buffered saline (PBS)” neutrophils, which are associated with cell survival regulation. (B) Graphic representation of neutrophil numbers normalized per liver tissue mass (in gram) in the injured liver in presence (PBS) and absence of Ly6C^hi monocytes and their MoMF descendants (MC-21), as analyzed by flow cytometry. (C) Flow cytometry image showing the discrimination between early apoptotic (AnnexinV⁺), late apoptotic (AnnexinV⁺PI⁺), and necrotic (PI⁺) neutrophils at 48 h following acetaminophen-induced liver injury (AILI). (D–F) Graphical summaries showing cell counts normalized per liver tissue mass (in gram) of (D) late apoptotic neutrophils, (E) necrotic neutrophils, and (F) early apoptotic neutrophils. (G) 3D-reconstructed confocal image generated from 20 µm Z-stacks of livers sections extracted at AILI-48 h from Cx3cr1^gfp/+ mice. Ly6C^hi monocyte-derived cells are in GFP and neutrophils are in red. Magnification ×63. Heat map analysis in A was performed on the differentially expressed genes (p < 0.05; t-test, ≥2-fold change). Data in (B–F) were analyzed by unpaired, two-tailed t-test, comparing at each time point livers from PBS (white) versus MC-21 (gray)-treated mice and are presented as mean ± SEM with significance: *p < 0.05, **p < 0.01, and ***p < 0.001 (n ≥ 5 mice/group for each time point).