FIGURE 2.

Bleomycin-induced acute lung injury. Mice received 33 µg bleomycin and were retrieved at 24 h. (A) Flow cytometric gating strategy to identify immune cell subsets and Gal-3 expression within the lung. After gating for single, live cells, leukocytes were selected based on CD45 expression. Alveolar macrophages (MΦ) (Siglec F⁺ CD11b^int CD11c⁺ CD64⁺), CD103⁺ dendritic cells (DCs - CD11c⁺ CD103⁺ CD24⁺), neutrophils (CD11b⁺ Ly-6G⁺) and eosinophils (SiglecF⁺ CD11b⁺ CD11c^–), followed by identification of the populations with overlapping expression patterns: interstitial macrophages (CD11b⁺ MHC II⁺ CD11c⁺ CD64⁺ CD24^–), CD11b⁺ DCs (CD11b⁺ MHC II⁺ CD11c⁺ CD24⁺ CD64^–), inflammatory monocytes (CD11b⁺ MHC II⁻CD64⁻ Ly-6C^hi) and patrolling monocytes (CD11b⁺ MHC II⁻CD64⁻ Ly-6C^low). (B) Interstitial leukocyte frequencies following acute bleomycin. (C) Galectin-3 expression in leukocyte subsets. Red line represents mean fluorescence intensity (MFI) in Gal-3 ^−/− mice. d) Gal-3 levels in serum and BAL. Data represented as mean ± SEM. Analysed via 1-way ANOVA (b,c) or students t-test. (D) (n = 6, *p < 0.05, **p < 0.01, ***p < 0.001).