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. 2021 Nov 4;12:6384. doi: 10.1038/s41467-021-26637-6

Fig. 4. Dysregulation of epithelial repair in α5SNP-expressing airway epithelium.

Fig. 4

a Scheme of lentiviral vectors (LV) used to locally express α5WT or α5SNP in tracheal epithelial cells in α5KO mice. b Top: GFP expression in tracheal epithelium. Middle and bottom: lymphocytes (CD45+) and macrophages (CD68+) in submucosa. Micrographs representative from three experiments. Scale bar: 100 μm. c Quantification of different epithelial phenotypes (normal epithelium, circles; basal cell hyperplasia, square; metaplasia, triangle; metaplasia/mild dysplasia, diamond-shaped) in mice with GFP LV (white symbols, n = 6), α5WT LV (gray symbols, n = 9) or α5SNP LV (blue symbols, n = 8) 12 days after lesion. One group of α5SNP (dark blue symbols, n = 8) received i.p. injections of Etanercept (α5SNP + Etan). Scale bar: 30 μm. d TNF-α and IL-1β production in GFP LV (white circles, n = 6), α5WT LV (gray circles, n = 9) or α5SNP LV (blue circles, n = 8) mice after lesion at D6, D9 and D12. e NF-κBiα expression in tracheal tissue extracts from α5WT or α5SNP LV mice on D6 after epithelial lesion with polidocanol (circles, n = 6 α5WT and n = 6 α5SNP LV mice) and pre-treatment with Etanercept (squares, n = 5 α5WT and n = 6 α5SNP LV mice). f Quantification of proliferation of tracheal epithelial cells isolated from GFP (n = 5), α5WT (n = 5) and α5SNP (n = 5) mice (O.D., optical density). g In vitro proliferation of epithelial cells isolated from α5WT or α5SNP LV mice (n = 6 per group) after exposure to TNF-α (squares, 10 and 50 ng/ml, 72 h). h Proliferative index quantification of human polyp epithelial cells isolated from WT (n = 18), HO (n = 21) or HT (n = 8) patients. For c–h, data are presented as mean ± SEM. Statistical analyses in (c) compared: WT vs GFP mice (§), α5SNP vs α5WT mice (*) and Etanercept treated vs non-treated α5SNP (#). *#,§p < 0.05; **, ##p < 0.01; ***p < 0.001 (Mann–Whitney two-sided test).