Fig. 2

Lung microenvironmental changes in response to osteosarcoma-secreted factors. A, B Heatmaps of transcripts encoding genes involved in ECM remodelling, inflammation, and immune cell recruitment in lungs from SCR-treated or PT-bearing mice compared with controls. C qRT-PCR analysis of S100A8/A9, Prg4, Cxcr2, Cxcl2, IL-1β, IL6, TGF-β, TNF-α, and FN1 genes in lungs from SCR-treated mice or PT-bearing mice compared with controls. (n = 3–7, per group). D Representative images of fibronectin, reticulin, α-SMA, and vimentin immunostaining at x200 magnification (Scale bar: 30 μm) in lung sections from CTR mice, SCR-treated mice, or carrying a PT. E Western blot analysis of fibronectin and collagen type IV. Expression levels with graphic quantification. F Schematic diagram illustrating the analysis of lung-infiltrating neutrophils and the peripheral blood in mouse models by flow cytometry. G Flow cytometric quantification of infiltrating neutrophils in the lungs and in the peripheral blood of control, SCR-treated and PT-bearing mice (n = 5–6, per group). Data are presented as mean ± SEM from 3–7 independent biological samples. *p < 0.05, **p < 0.01 compared to control lungs (Mann-Whitney test)