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. 2021 Jul 13;54(7):1494–1510.e7. doi: 10.1016/j.immuni.2021.04.025

Figure 5.

Figure 5

rTEM neutrophils stemming from locally injured aged tissues accumulate in the lungs.

Young and aged mice were subjected to sham or cremasteric IR injury.

(A) Representative confocal images of post-capillary venules (PCVs) immunostained for CD31 and MRP14 (neutrophils) in WT mice (scale bar: 20 μm).

(B) Representative confocal images and quantification of lung vascular leakage in WT mice 4 h post reperfusion (scale bar: 20 μm; n = 4-5 mice/group).

(C) Neutrophil normal TEM events and (D) frequency of neutrophil rTEM in Y→Y or Y→A chimeras (see Figure 1H) as assessed by confocal IVM (n = 6 mice/group).

(E-I) Mice were injected i.v. with a biotinylated anti-Ly6G mAb and AF647-Strept locally applied to the cremaster muscle.

(E) Time-lapse confocal IVM images (Video S4) of a neutrophil rTEM event in an aged Lyz2-EGFP-ki cremaster muscle during IR injury illustrating that the neutrophil exhibiting rTEM is AF647-Strepthi (Top panel: en face luminal view; bottom panel: isolated neutrophil; scale bar: 4 μm).

(F-I) Representative flow cytometry profiles and frequency of AF647-Strepthi neutrophils in (F-G) blood and (H-I) pulmonary vascular washouts in WT mice (n = 4-11 mice/group). Numbers indicate the percentage of AF647-Strepthi neutrophils. Means ± SEM, #p < 0.05, ###p < 0.001, ####p < 0.0001 as compared to age-matched controls, p < 0.05, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001 as indicated.

See also Figure S5.