Fig. 3

Frequencies of TIE2-negative and -positive monocytes and macrophages during murine ALI. Female C57BL/6 mice (10–12 weeks old) were challenged by intratracheal instillation of LPS (5 μg/g bodyweight), and 24 or 72 h thereafter, animals were sacrificed and frequencies of TIE2-negative and -positive monocytes and macrophages were determined by FACS analysis in blood, spleen, bronchoalveolar lavage (BAL) fluid, and the lungs. a Frequencies of monocytes and macrophages (expressed as percentage of all counted cells) are shown on the left side, and fractions of TIE2-expressing cells (expressed as percentage of all monocytes or macrophages, respectively) are shown on the right side. Mean ± SD, n = 7, Mann-Whitney U test, * p < 0.05 (vs. PBS control). b Representative FACS dot plots of lungs from control and ALI animals, respectively, sacrificed 24 or 72 h following ALI induction. Analysis of neutrophil granulocytes (NG) and macrophages (MPh) is shown on the left side, and that of the TIE2-expressing macrophage fraction (TEM) is shown on the right side. c Comparison of frequencies of TIE2-expressing cells (expressed as percentage of all monocytes or macrophages, respectively) in all four analyzed compartments. Mean ± SD, n = 7, Kruskal-Wallis test with Dunn‘s posthoc analysis, *p < 0.05 (vs. blood, vs. spleen), § p < 0.05 (vs. blood), # p < 0.05 (vs. blood, vs. BAL). d Ratio between relative frequencies of TIE2-positive and -negative monocytes/macrophages cells in spleen (left) and lung tissue (right). Mean ± SD, n = 7, Mann-Whitney U test, * p < 0.05 (vs. 24 h control, vs. 72 h ALI), # p < 0.05 (vs. 72 h control, vs. 24 h ALI). e TIE2 expression in THP-1 cells after incubation with LPS or recombinant human TNF-α (concentration as indicated in μg/ml) for 24 h, respectively. Mean from biological triplicates ±SD, Kruskal-Wallis test with Dunn‘s posthoc analysis, * p < 0.05 (vs. unstimulated control). ALI = Acute Lung Injury, BALF = BAL fluid