Figure 1. Phenotype and kinetics of aged neutrophils in blood.

(A) Flow cytometric analysis of CD62L and CXCR4 expression in Ly6G⁺ blood neutrophils. n=5 mice.

(B) Number of CD62L^LO neutrophils in the blood of WT, PEdKO and Fut7−/− mice. n=8–18.

(C) Scheme of transfer experiments of CD62L^LO CD45.2⁺ and CD62L^HI CD45.1⁺ neutrophils into GFP⁺ mice. Graphs show the absolute number of CD62L^HI or CD62L^LO neutrophils derived from donor cells at different times after transfer. Insets show the relative changes of each population. n=5 mice. Statistics are vs. t = 5 min.

(D) Intensity of CXCR4 expression in CD62L^HI and CD62L^LO neutrophils 5 and 480 min after transfer. n=5 mice.

(E) Density plots and frequencies of CD62L^HI and CD62L^LO neutrophils among Ly6G⁺ BrdU⁺ cells at different times post-labeling. n=5 mice. Statistics are vs. 48h. See also Figure S1B.

(F) Bright-field micrographs of sorted CD62L^HI and CD62L^LO blood neutrophils (scale bar, 10 μm), and frequency of hypersegmented neutrophils in blood. n=3–4 mice.

(G) Cytometry plots showing CD62L and CXCR4 expression in blood neutrophils at different times of the day. Indicated also are the mean percentage ± s.e.m. of CD62L^LO cells out of total neutrophils at each time. The right graph shows the number of total, CD62L^HI and CD62L^LO neutrophils at the same times. Colored boxes highlight periods of clearance and release. n=5–9 mice. Statistics are vs. ZT5. Data are shown as mean ± s.e.m. *P<0.05, **P<0.01, ***P<0.001. See also Figure S1.