Fig. 6: Macrophages promote fibroblast activation via the IL6-STAT3 axis.

a GSEA analysis showing TOP5 enriched hallmark pathways in Granulation group compared with Normal group. b GSEA analysis show IL6 jak STAT3 pathway enrichment in Granulation group. c Box Plot showing the JAK-STAT pathway score in all clusters in Normal group and Granulation group: B cells (n = 1434), basal cells(n = 10982), ciliated cells (n = 1317), endothelial cells (n = 7241), epithelial cells (n = 5123), fibroblasts (n = 6808), mast cells (n = 475), macrophages (n = 5625), neutrophils (n = 10178), plasma cells (n = 1249), proliferating cells (n = 824), SMCs (n = 3608), T cells (n = 3500) and Type II alveolar cells (n = 222). n represents the number of cells in cluster. d Representative images of P-STAT3 and P-STING immunohistochemical staining. (n = 3 sample per group). e Heatmap illustrates ligand-receptor interactions between Macrophages, T cells, neutrophils, and fibroblasts. f–r Mice were divided into three groups: control, BAS + PBS and BAS + Clodronate Liposomes. f Schematic of Clodronate Liposomes administration in mice. g Representative images of H&E staining in different groups at days 7 and 14. h Quantitative analysis of LP thickness in different groups. (n = 4 mice per group). i Micro CT scans of different groups in Horizontal, Coronal and Sagittal plane. j SYNPASE 3D reconstruction of mouse trachea. Red arrows indicating the stenosis site in (i–k). The CT diagram of the area of tracheal stenosis measured by SYNPASE 3D and quantitative analysis of area in different groups(n = 4 mice per group). l Representative images of Masson staining in different groups. m Quantitative analysis of collagen area in different groups (n = 4 mice per group). n, o Representative tracheal immunofluorescence images and mean fluorescence intensity of αSMA(red) and COL1(green) in different groups (n = 3 mice per group). p GSEA analysis show IL6 jak STAT3 pathway enrichment in BAS + PBS group. q, r Representative tracheal immunofluorescence images and mean fluorescence intensity of P-STAT3(green) and IL6(red) in different groups (n = 3 mice per group). Scale bars in (d, g, n, q) indicates 200 μm and 50 μm. Scale bars in (l) indicate 50 μm. In (c), box bounds shows 25th and 75th percentiles, whiskers shows 25th percentiles minus 1.5*IQR to 75th percentiles plus 1.5*IQR and box center shows the median. Data are presented as the mean ± SEM. One-way ANOVA analysis followed by Tukey post hoc multi-comparison test was used in (h, k, m, o, r). One-way ANOVA analysis followed by Duncan multiple- range test was used in c. Source data are provided as a Source Data file.