Leukocyte recruitment and vascular permeability, 5 nmol/L MIP-2 (A–C) and 0.5 nmol/L MIP-2 (D–F) superfusion.

<p>Leukocyte adhesion (A and D), emigration (B and E) and microvascular permeability increase (C and F) before (time 0) and after addition of MIP-2 (5 nmol/L in A–C and 0.5 nmol/L in D–F) in the superfusate in WT (n = 5 for each concentration) and Mac-1-/- mice (n = 5 for each concentration). Untreated wild-type mice (n = 5) are also included. A leukocyte was considered to be adherent if it remained stationary for more than 30 s, and was quantified as the number of adherent cells within a 100 µm length of venule during 5 min. Leukocyte migration was defined as the number of cells in the extra vascular space within a 200×300 µm area. All values are means±SE. * p<0.05 compared to C57Bl/6 mice receiving similar treatment at the same time-point.</p

Endothelial enscapsulation of transmigrating neutrophils.

<p>Electron micrographs and cartoons of transmigrating wild-type (A) and Mac-1 deficient neutrophils (B). The arrowheads mark the junctions seen in A, the thin endothelial sheet that covers the transmigrating cells is marked with *, e1, e2 and n represent individual endothelial cells and neutrophils respectively. The arrows in B mark gaps in the endothelium, and en marks the endothelial nucleus. Scale bars correspond to 1 µm. The images represent 1 out of 40 analyzed wild-type and 1 out of 24 Mac-1-/- neutrophils. In panel C are presented the percentages of the adherent neutrophils that were covered by endothelial dome-structures.</p

Endothelial dome formation using spinning disk confocal microscopy.

<p>Spinning disk confocal images of MIP-2 superfused cremaster muscle of mice with GFP positive endothelium (green) and GR-1 Alexa Fluor 555 stained neutrophils (red). Panels A and B show adherent neutrophils that have been covered by endothelium. By following the same vessel with two adherent neutrophils over time, dynamic formation of endothelial domes was revealed.</p

Transmigration in respect to endothelial junctions.

<p>The distribution of transmigrating wild-type (n = 40) and Mac-1-/- neutrophils (n = 24) in respect to junctions on electron micrographs.</p

Endothelial transmigratory cup formation.

<p>Electron micrographs and cartoons of transmigrating wild-type neutrophils where the endothelium forms a transmigratory cup (A, B and C). The arrowheads mark the junctions and L marks the lumen of the blood vessel. e1, e2 and n represent individual endothelial cells and neutrophils respectively. The bar corresponds to 1 µm.</p