Figure 1
From: Activation of C3a receptor is required in cigarette smoke-mediated emphysema
Complement 3 is required in smoke-induced emphysema development. WT or C3−/− mice were exposed to cigarette smoke or air for 6 months. (a) Representative images of H&E-stained lung sections from WT and C3−/− mice exposed to air or cigarette smoke. Representative of three independent studies (n=4–5 per group). (b) Micro-CT quantification of lung volume and (c) MLI using unbiased morphometry in the indicated groups of mice (n=10). *P<0.05, ***P<0.001, as determined by student t test and one-way ANOVA with Bonferroni’s multiple comparison. (d) BAL fluid analyses from the same group of mice (n=4 or 5 per group) showing macrophages (Mac), lymphocytes (Lymph), and neutrophils (Neu). **P<0.01, as determined by one-way ANOVA with Bonferroni’s multiple comparison. Expression of Mmp9 (e) and (f) Mmp12 mRNA in BAL cells was measured by qPCR. **P<0.01, as determined by the one-way ANOVA with Bonferroni’s multiple comparison. (g) Representative and (h) cumulative flow cytometry analysis of B220−CD11b+CD11c+ mDCs in the lung of the same group of mice (numbers in each quadrant indicate % positive cells for the indicated cytokines); right panel cumulative data (n=4 or 5 mice in each group). **P<0.01, as determined by the one-way ANOVA with Bonferroni’s comparison. Cumulative intracellular cytokine staining of IL-17A in αβ (i), and γδ (j) CD3+/CD4+T cells (n=4–5 in each group). *P<0.05, as determined by the one-way ANOVA with Bonferroni’s comparison. (k) and (l) Expression of Il6 and Il1beta mRNA in BAL cells were measured by qPCR. **P<0.01, ***P<0.001, as determined by the one-way ANOVA with Bonferroni’s comparison. (m) Expression of Cd86 mRNA in BAL cells was measured by qPCR. **P<0.01, as determined by the one-way ANOVA with Bonferroni’s multiple comparison. IL-6 concentration (n) and (o) TGFβ concentration in mouse lung homogenate and (p) KC level in BAL fluid was measured by multiplex assay and ELISA (TGFβ). *P<0.05, as determined by the one-way ANOVA with Bonferroni’s comparison. All mRNA gene expression was normalized to 18S expression and analyzed by ΔΔCt method. Results are represented as mean±s.e.m. from three independent experiments with 4–5 mice in each group. ANOVA, analysis of variance; H&E, hematoxylin and eosin; mDCs, myeloid dendritic cells; micro-CT, micro-computed tomography; MLI, mean linear intercept; qPCR, quantitative reverse transcription PCR; IL, interleukin; WT, wild type. Scale bar: 100 μm.
