Fig. 4From: Differential effects of Nintedanib and Pirfenidone on lung alveolar epithelial cell function in ex vivo murine and human lung tissue cultures of pulmonary fibrosisEx 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 < 0.05Back to article page