Fig. 7. Epithelial depletion of CAR induces TGF-β-dependent airway remodelling.

a Representative images of sections of lungs from mice under specified conditions stained for fibronectin (antibody; brown) and collagen (Sirius red; pink) staining. Scale bar: 100 mm. b Quantification of fibronectin and collagen staining around the airways from 20 airways in FFPE lung sections from each experimental group; from 5 mice per condition (one lung per mouse) representative of 3 independent experiments. c Quantification of proliferation of primary human lung fibroblasts and airway smooth muscle cells (SMC) treated with supernatants from 16HBE parental or CAR CRISPR cells for 48 h. Data. d Lysates from primary human lung fibroblasts treated as in c and probed for fibronectin and collagen I. e Quantification of 4 independent experiments from data as in d. f TGF-β levels measured by ELISA in supernatants from 16HBE parental and CAR CRISPR cells. 3 replicates per condition, pooled from 3 independent experiments. g TGF-β transcript levels measured by qPCR in 16HBE parental and CAR CRISPR cells. Data shown is pooled from 5 independent experiments. h Quantification of proliferation of primary human airway SMC treated with supernatants from 16HBE parental or CAR CRISPR cells for 48 h in the presence or absence of a TGF-β inhibitor (R26). Pooled from 3 independent experiments with 3 replicates per condition per experiment. i Lysates from cells as in h probed for collagen I and HSC70. Representative of 3 independent experiments. All graphs show median (line) with 25/75 percentiles (box) and min/max values. Unpaired 2-tail student’s T-tests were performed on data in c, e, f, g; One-way ANOVA analysis with Dunnett’s post hoc test was performed to test statistical significance in b; two-way ANOVA analysis with Tukey’s post hoc test was used for h. P values *p < 0.05; **p < 0.01; ***p < 0.0005. Source data are provided as a Source Data file.