Loss of PHD2 in neutrophils enhances the speed of transendothelial migration in a mouse model of acute skin inflammation. Schematic representation of the acute skin inflammation model (A), including time points for the intravital microscopy analysis (B) and final histological analyses (D-G). (B-C) Intravital imaging of representative neutrophils using 2-photon microscopy, 3 hours after PMA was applied to the ear (WT vs cKO P2). Time point 00m00s shows Ly6G⁺ neutrophils (+ white dashed lines surrounding the red cell) that had stopped rolling/moving at the blood-endothelial barrier (= white dashed lines) before diapedesis and migration into the inflamed tissue. The last frames represent the first time point that the individual neutrophil had completely left the bloodstream (scale bars, 10 μm). (C) The average time necessary for individual neutrophils to complete the diapedesis. Total amount of cells were collected from 3 individual mice per genotype. Representative immunofluorescent images of Gr1⁺ neutrophils on ear sections 24 hours after PMA application (D-E, scale bars, 50 μm) and quantification of the total number of cells per area or fraction of apoptotic neutrophils (cCas3⁺) (F-G). Each data point represents an average amount based on ≥6 images per individual mouse. (H-I) Extracellular acidification rate (ECAR) measurements from steady-state neutrophils immediately after negative selection. Data points represent individual mice from ≥3 experiments - normalized values against WT control. Data are mean ± standard error of the mean. *P < .05, Mann-Whitney U test.