Fig. 5: High-affinity LFA-1 is localized at the trailing edge of the neutrophil during the elongation step of extravasation upon stimulation with N-formylmethionyl-leucyl-phenylalanine. | Experimental & Molecular Medicine

Fig. 5: High-affinity LFA-1 is localized at the trailing edge of the neutrophil during the elongation step of extravasation upon stimulation with N-formylmethionyl-leucyl-phenylalanine.

From: LFA-1 (CD11a/CD18) and Mac-1 (CD11b/CD18) distinctly regulate neutrophil extravasation through hotspots I and II

Fig. 5

a CD18 expression (upper left panel), CD11a expression (upper right panel), Alexa 594-anti-CD31 antibody staining of an endothelial cellular border, and the CD18-to-CD11a ratio (lower right panel), visualized in the step of uropod elongation of a neutrophil in LFA-1 FRET (CD11a-mYFP/CD18-mCFP) mice. Two-photon intravital imaging of LFA-1 FRET mice was performed. The cyan fluorescent protein/yellow fluorescent protein (CFP/YFP) ratio is visualized using the rainbow scale. The arrow indicates LFA-1 affinity status at the trailing edge of an elongated neutrophil. The dotted arrow indicates the elongation of an extravasating neutrophil. Scale bar, 10 μm. See Video 10. The images are representative of at least five independent videos. b CD18 expression (upper left panel), CD11b expression (upper right panel), Alexa 594-anti-CD31 antibody staining of an endothelial cellular border, and the CD18-to-CD11a ratio (lower right panel), visualized in the step of uropod elongation of a neutrophil in Mac-1 FRET (CD11b-mYFP/CD18-mCFP) mice. Two-photon intravital imaging of Mac-1 FRET mice was performed. The CFP/YFP ratio was visualized using the rainbow scale. The arrow indicates the Mac-1 affinity status at the trailing edge of an elongated neutrophil. The dotted arrow indicates the elongation of an extravasating neutrophil. Scale bar, 10 μm. See Video 11. The images are representative of at least five independent videos. c The affinities of LFA-1 and Mac-1 were compared at the trailing edges of neutrophils during uropod elongation. *P < 0.0001. d Plot of the correlation of cell length with LFA-1 affinity. Data from two-photon intravital imaging of neutrophils in LFA-1 FRET mice during uropod elongation, as the final step of neutrophil extravasation, were analyzed. A dot indicates the correlation value of an elongated neutrophil. e Plot of the correlation of cell length with Mac-1 affinity. Data from two-photon intravital imaging of neutrophils in Mac-1 FRET mice during uropod elongation, as the final step of neutrophil extravasation, were analyzed. A dot indicates the correlation value of an elongated neutrophil

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