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. 2020 Feb 7;11:68. doi: 10.3389/fimmu.2020.00068

Figure 5.

Figure 5

Supplementation with Del-1 attenuates the pathological characteristics of PF in mice. (A) Representative ex vivo micro-computed tomography (μ-CT) scans of lung sections from WT and Del-1−/− mice with BLM (1 U/kg)-induced PF, which were supplemented intravenously with bovine serum albumin (BSA; 50 μg/dose) or soluble Del-1 (sDel-1; 50 μg/dose) at 5, 9, and 13 days post-BLM administration (dpa). Lung sections were taken at 21 dpa. Dashed lines indicate fibrotic regions. Quantification of fibrosis in the μ-CT images is shown in the right panel. Data are represented as the mean ± SEM (n = 3 mice per group). *p < 0.05; n.s., not significant; Mann-Whitney U-test. (B) Survival analysis of WT and Del-1−/− mice with BLM-induced PF (1.5 U/kg BLM) that were supplemented with BSA or sDel-1 (intravenous injection, 50 μg/dose sDel-1 at 5, 9, and 13 dpa [n = 12 mice per group]). *p < 0.0001; log-rank test. (C) The levels of active TGF-β and LAP in the lungs of mice with BLM-induced PF that were supplemented with Fc or Del-1-Fc (intravenous injection; 50 μg/dose at 5, 9, and 13 dpa). The lung lysates were collected at 14 dpa and subject to ELISA. Data are expressed as mean ± SEM (n = 3 mice per group). *p < 0.05, ***p < 0.001; Mann-Whitney U-test. (D) Representative western blots to detect the expression of α-SMA and the level of phosphorylated Smad3 in WT mice with BLM-induced PF (2 U/kg BLM) supplemented with Fc or Del-1-Fc (intravenous injection; 50 μg/dose Fc or Del-1-Fc at 14, and 18 dpa). Lung lysates were collected at 21 dpa. Densitometric data are shown in the right panels. Data are expressed as mean ± SEM (n = 3–4 mice per group). **p < 0.01, ***p < 0.001; Student's t-test. (E) Hydroxyproline analysis of lung tissues from WT mice with BLM-induced PF (2 U/kg BLM) supplemented with Fc or Del-1-Fc, as in (D). Data are expressed as the mean ± SEM (n = 3–4 mice per group). *p < 0.05, ***p < 0.001; Student's t-test. (F) Hydroxyproline analysis of lung and skin tissues from WT mice with systemic fibrosis (8 U/kg/dose BLM, daily subcutaneous injection for 15 days) supplemented with Fc or Del-1-Fc (intravenous injection; 50 μg/dose Del-1-Fc at 7, 11, 15, and 19 dpa after the first BLM instillation). Tissues were harvested at 23 days after the first BLM instillation. Data are expressed as the mean ± SEM (n = 3 mice per group). *p < 0.05; Mann-Whitney U-test.