Fig. 7: Ablation of FOXN3 S412 and S416 phosphorylation impends pulmonary fibrosis.

a A Western blot assay was performed to assess the alterations in FOXN3 and Smad4 expression levels in the lungs of WT or Foxn3^KI/KI mice harboring double mutations at S379 and S383 (n = 3). b Histological trichrome staining analysis comparing the fibrotic response in lungs of WT and Foxn3^KI/KI mice, with or without BLM stimulation. Scale bars, 100 μm. c H&E staining analysis showing the inflammatory response in lungs of WT and Foxn3^KI/KI mice, with or without BLM stimulation. Scale bars, 100 μm. d, e ELISA analysis was performed to detect the levels of TNFα (d) and IL-6 (e) in the BAL fluid obtained from the lungs of BLM-induced WT or Foxn3^KI/KI mice (n = 4). f A quantitative PCR assay showing the RNA levels of pro-fibrotic factors in lungs of WT and Foxn3^KI/KI mice, with or without BLM stimulation (n = 4). g A hydroxyproline assessment was conducted to evaluate the collagen content in the lungs of WT and Foxn3^KI/KI mice, with or without BLM stimulation (n = 4). h Quantitative PCR analysis was performed to examine the impact of FOXN3 phosphorylation on the expression levels of pro-fibrotic factors in primary pulmonary fibroblasts isolated from WT and Foxn3^KI/KI mice following BLM treatment (n = 3). The data (d, e, and h were assessed by two-tailed Student’s t test, and the data (f and g) were assessed by one-way ANOVA. All data are shown as the mean ± SD.