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. 2020 Dec;190(12):2355–2375. doi: 10.1016/j.ajpath.2020.08.018

Figure 8.

Figure 8

Fewer macrophages and less fibrosis after bleomycin challenge in lungs with expanded lymphatics. A–C: Staining of macrophages for allograft inflammatory factor 1/ionized calcium binding adapter molecule 1 (Aif1/Iba1). D: Counts of macrophages in hot spots in lung parenchyma after bleomycin administration. Macrophage numbers significantly increased in hot spots after bleomycin treatment, but significantly less in CCSP/VEGF-C lungs than in control lungs. E–H: Low-magnification panoramas of left lung of C57BL/6 mice stained for type I collagen, showing weak staining at baseline (E). After bleomycin challenge, staining is weak at 7 days (F), patchy at 14 days (G), and uniformly strong at 28 days (H). I: Measurements of type I collagen staining plotted as cumulative frequency of immunofluorescence pixel intensities in images at baseline and 7, 14, or 28 days after bleomycin challenge (E–H). Curves for 14 and 28 days significantly different from baseline. J: Relative amounts of type I collagen shown by height of curves plotted from group-related differences in cumulative frequencies from I. Taller curves indicate larger differences. K: Amounts of type I collagen in lungs of groups in I and J compared by area density measurements at threshold = 30 based on maximal difference in curves. Dotted line: zero on y axis. L: Less type I collagen staining at 28 days after bleomycin challenge in the lung of CCSP/VEGF-C mouse with expanded lymphatic network than in control mouse (H). M: Measurements of type I collagen staining in lungs as in K, comparing large increase at 28 days after bleomycin challenge in control mice with a significantly smaller increase in CCSP/VEGF-C mice. Dotted line: zero on y axis. N: Measurements of hydroxyproline in right lungs, showing increase over baseline group at 28 days after bleomycin challenge. Dots represent individual mice. Blue dots, baseline (no treatment). However, no significant difference was found between controls (black dots) and CCSP/VEGF-C mice (red dots). n = 5 to 7 mice per group (D); n = 4 to 7 mice per group (J); n = 5 to 12 mice per group (N). ∗P < 0.05 versus baseline [analysis of variance (ANOVA)]; †P < 0.05 versus control (ANOVA). Scale bars: 50 μm (A–C); 1 mm (E–H and L).