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. 2019 Oct 14;129(11):4962–4978. doi: 10.1172/JCI127959

Figure 3. GGOH-mediated lung fibrosis occurs in the absence of injury.

Figure 3

Ten days after exposure of WT mice to saline or bleomycin, pumps containing vehicle or GGOH were s.c. implanted; mice were sacrificed 11 days later. (A) Caspase-3 activity in AECs (n = 5/group). (B) Representative images of lung IHC for SPC (green), TUNEL (red), and DAPI (blue) (n = 4 per group) and (C) quantification of TUNEL+ cells from B (n = 4/group). Scale bars: 20 μm. WT mice were exposed to saline or bleomycin. Confocal imaging and total number of BAL cells stained with (D and E) CD11c (n = 5–9) and (F and G) CD11b (n = 5–9), together with DAPI. Scale bars: 50 μm. Ten days after exposure of WT mice to saline or bleomycin (Bleo), pumps containing vehicle or GGOH were s.c. implanted, and lungs were extracted 11 days later. (H) Representative flow cytometric plots of MDMs (CD45+CD11b+/–Ly6G–CD64+Ly6c–Siglec Flo) and RAMs (CD45+CD11b+/–Ly6G–CD64+Ly6c–Siglec Fhi). FSC-A, forward scatter area. Total numbers of (I) MDMs and (J) RAMs from BAL. WT or CCR2–/– mice were exposed and treated as described above. (K) Rac1 activity (n = 4–5) and mRNA expression of (L) Tgfb1 (n = 4–5), (M) Arg1 (n = 4–5), (N) Tnf (n = 4–5), and (O) Nos2 (n = 4–5). Inset in K shows CCR2 immunoblot analysis. Values indicate the mean ± SEM. **P < 0.001 and ***P < 0.0001, by 2-tailed Student’s t test (E and G) and 1-way ANOVA followed by Tukey’s multiple comparisons test (A, C, and I–O).