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. 2021 Jun 9;7(24):eabg6005. doi: 10.1126/sciadv.abg6005

Fig. 8. Ghr vesicles reduce pulmonary fibrosis in mesenchymal Ghr-deficient mice.

Fig. 8

(A) Immunostaining of red fluorescent protein, Scgb1a1, Sftpc, and DAPI in mouse lungs administrated with mCherry sequence containing Ghr vesicles. Yellow solid line, mCherry and Scgb1a1 or Sftpc colocalized cells; white dotted line, Scgb1a1- or Sftpc-negative cells. (B) Ghr mRNA expression in ATII cells sorted from mouse lungs treated with Ghr or control vesicles. (C) Ghr protein expression in ATII cells sorted from mouse lungs treated with Ghr or control vesicles. (D) A schematic diagram depicting the administration of Ghr or control vesicles into the lungs of GhrΔBMC mice after bleomycin treatment. Mouse lungs were harvested on day 21. (E) Significantly decreased hydroxyproline levels were detected in Ghr vesicles and control vesicle–administered GhrΔBMC mouse lungs (Ctrl, n = 18; GhrΔBMC + Ctrl vesicle, n = 16; GhrΔBMC + Ghr vesicle, n = 18). (F) Cxcl1, Cxcl2, Cxcl5, Cxcl10, Cxcl11, Cxcl12, and Cxcl16 protein secretion in BALF of GhrΔBMC mice with Ghr or control vesicle treatments. (G and H) Representative images of H&E staining (G) and trichrome staining (H) in Ghr or control vesicle–treated GhrΔBMC mouse lung 21 days after bleomycin injury. (I) A diagram illustrating how subepithelial mesenchymal cells support ATII cells through Ghr vesicle and Ghr-coordinated CXC chemokines in normal and fibrotic conditions in the lung. Scale bars, 10 μm (A), 100 μm (G and H, higher power), and 1 mm (G and H, lower power). *P < 0.05, **P < 0.005, ***P < 0.001, and ****P < 0.0001 by unpaired two-tailed Student’s t test (B and C) and one-way ANOVA (E), means ± SEM.