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. 2025 Jul 25;15(16):8360–8376. doi: 10.7150/thno.117523

Figure 4.

Figure 4

PTEN knockout alleviates lung injury and epithelial mesenchymal transition (EMT) in vivo. (A) Schematic representation of the experimental procedure. PTENCKO mice were induced with pulmonary fibrosis using HCl and mechanical ventilation (MV). (B) Western blot analysis of PTEN protein expression in lung tissue from PTENF/F and PTENCKO mice. GAPDH served as a loading control. (C) Quantification of PTEN expression levels normalized to GAPDH. Data are presented as mean ± SEM. n = 4 mice/group. (D) Immunofluorescence staining for PTEN (green) in lung sections from PTENF/F and PTENCKO mice. Nuclei were counterstained with DAPI (blue). (E) Hematoxylin and Eosin (HE) and Masson's trichrome staining of lung sections showing reduced tissue damage and collagen deposition in PTENCKO mice compared to PTENF/F mice after HCl treatment with or without MV. (F) Quantification of Ashcroft fibrosis score in PTENCKO mice compared to PTENF/F mice after HCl treatment with or without MV. Data are presented as mean ± SEM. n = 4 mice/group. (G-I) Quantification of Collagen-I, hydroxyproline in lung tissue, and TGF-β1 levels in bronchoalveolar lavage fluid (BALF). Data are presented as mean ± SEM. n = 4 mice/group. (J) Immunofluorescence staining for E-cadherin (red), Vimentin (green), and α-SMA (green) in lung sections from PTENCKO and PTENF/F mice under HCl + MV condition. Nuclei were counterstained with DAPI (blue). (K) Western blot analysis of E-cadherin, Vimentin, α-SMA protein expression in lung tissue from PTENCKO and PTENF/F mice under different treatment conditions. (L-N) Quantification of E-cadherin, Vimentin, α-SMA expression levels normalized to GAPDH in lung tissue from PTENCKO and PTENF/F mice under different treatment conditions. Data are presented as mean ± SEM. n = 4 mice/group. Comparisons between two groups were performed using independent sample t tests. One-way ANOVA and the Bonferroni post hoc test were used to compare more than two groups. Not significant (ns): P > 0.05, *: P < 0.05, **: P < 0.01, ***: P < 0.001, ****: P < 0.0001.