Fig. 3

Increased deposition of fibronectin and collagen by activated lung fibroblasts favour the adhesion of OS cells to the lung ECM. A Schematic diagram of the establishment of the primary cultures of fibroblasts isolated from the lungs of untreated (NFs), SCR-treated (NAFs^SCR) and PT-bearing mice (NAFs^PT). B Representative image of immunofluorescence staining for α-SMA, FAP, vimentin, fibronectin and phalloidin at x20 magnification (Scale bar: 50 μm) in primary cultures of fibroblasts from CTR, SCR-treated or PT-bearing mice. Immunofluorescence quantification of α-SMA, FAP, vimentin and fibronectin (n = 3, per group). C Relative adhesion of 143B cells to increasing concentrations of fibronectin and collagen type IV ranging from 1 to 10 µg/mL (n = 3–4, performed in triplicate). Representative bioluminescence images of cell adhesion (143B cells) to different concentrations of fibronectin and collagen IV. D Representative SEM images (Scale bar: 50 μm) of decellularized lung sections and fibronectin and collagen immunostaining at x20 magnification (Scale bar: 50 μm) in decellularized lung sections from CTR mice, SCR-treated mice, or carrying a PT. E Relative adhesion of 143B cells to decellularized lung sections from CTR, SCR-treated, or PT-bearing mice (n = 5). Representative pictures of the decellularized fragments in the wells (upper row) and bioluminescent images of adhered cells (lower row). The bioluminescent signal is represented as radiance (p/s/cm²/sr). Data are presented as mean ± SEM. *p < 0.05 and ****p < 0.0001 are significantly different when compared to NFs (Kruskal-Wallis (B)); *p < 0.05 and **p < 0.01 when compared to healthy decellularized lungs (One-way ANOVA (E))