Redistribution of neutrophils around wounds. (a) Experimental scheme of procedure for the wound healing model in the ear. (b) Representative image for neutrophil recruitment to the ear wound 24 h after skin injury in Ly6G^tdTomato mice showing neutrophils (red) and blood vessels (cyan). Dotted black lines represent the surrounded neutrophils. (c) Neutrophil numbers in skin wounds of WT mice injured at ZT5 and ZT13, and ATI2341-treated mice, quantified by flow cytometry; n = 6–7 mice per group. (d) Whole-mount representative images of neutrophil distribution (left) in and around skin wounds of vehicle- and ATI2341-treated CXCR4^ΔN mice injured at ZT5, quantified in bars at right; n = 6 mice per group (1–2 wounds per mouse). Solid white lines limit the neutrophil ring and dotted white lines the adjacent area to the wound. (e) Representative images (left) and quantification (right) of Annexin V signal around skin wounds from neutropenic (Mcl1^fl/fl) and control mice; n = 4–6 mice per group (2–5 wounds per mouse). (f) Representative confocal images (left) and quantification (right) of TUNEL staining for identification of apoptotic areas around skin wounds in control mice (ZT5), mice injured at night (ZT13), and ATI2341-treated mice; n = 3 mice per group (1–2 wounds per mouse). Data in c and d are pooled from two experiments. Data in e are pooled from four experiments. Data in f are from a single experiment. Data are shown as the mean ± SEM. *P < 0.05; ns, not significant, as determined by one-way ANOVA (c and f) and unpaired t test (d and e).