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Fig. 4 | Respiratory Research

Fig. 4

From: Differential effects of Nintedanib and Pirfenidone on lung alveolar epithelial cell function in ex vivo murine and human lung tissue cultures of pulmonary fibrosis

Fig. 4

Ex vivo treatment with Nintedanib stimulates alveolar epithelial marker expression in the human 3D-LTCs model of early pulmonary fibrosis. a Schematic of treatment with fibrotic cocktail (FC) or control cocktail (CC) and Nintedanib (Nint) and downstream analysis. 3D-LTCs were generated and treated with FC or CC for 48 h before FC or CC treatment was replenished and Nintedanib or control treatment was added. Treatment was stopped after 120 h and downstream experiments were performed. b Representative Immunofluorescence of punches treated with CC or FC for 120 h and stained for Fibronectin. Scale bars represent 1 mm. c-f Punches were treated with CC/FC and Nintedanib (1 μM) as indicated in (a). c Metabolic activity of punches 120 h after treatment with CC/FC and co-treatment with Nintedanib. N = 3. Significance was assessed by two-way ANOVA followed by Sidak’s multiple comparisons test. d Gene expression analysis by qPCR of epithelial cell marker SFTPC, NKX2.1, CDH-1, ZO-1. Log fold change is presented as mean ± SEM, N = 3. Means were compared to respective DMSO control using one-sample t-tests in comparison to a hypothetical value of 0. e Representative Immunofluorescence of punches treated with FC and Nintedanib for proSP-C. Scale bars represent 140um. f SP-C secretion of punches 120 h after treatment with CC/FC and co-treatment with Nintedanib was measured by ELISA. Values shown are normalized to CC treatment. Significance was assessed using Wilcoxon matched pairs test. N = 6. Significance: *p < 0.05

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