Figure 7.

PTEN regulates epithelial mesenchymal transition (EMT) via P53-senescence axis. (A) Schematic representation of the experimental design involving P53 activation using the MDM2-P53 complex inhibitor RG7388 in PTEN^CKO or PTEN^F/F mice subjected to HCl and mechanical ventilation (MV). (B) Hematoxylin and Eosin (HE) and Masson's trichrome staining of lung sections from PTEN^F/F, PTEN^CKO, and PTEN^CKO + RG7388 mice under HCl+MV condition. (C) Quantification of Ashcroft fibrosis score in PTEN^F/F, PTEN^CKO, and PTEN^CKO + RG7388 mice under HCl+MV condition. Data are presented as mean ± SEM. n = 4 mice/group. (D-F) Quantification of Collagen-I, hydroxyproline in lung tissue, and TGF-β1 levels in bronchoalveolar lavage fluid (BALF) under HCl+MV condition. Data are presented as mean ± SEM. n = 4 mice/group. (G) Western blot analysis of P53 and P21 protein expression in lung tissue from PTEN^F/F, PTEN^CKO, and PTEN^CKO + RG7388 mice under HCl+MV condition. (H-I) Quantification of P21 and P53 expression levels normalized to GAPDH. Data are presented as mean ± SEM. n = 4 mice/group. (J) Immunofluorescence staining for P53 (red) and P21 (red) in lung sections from PTEN^F/F, PTEN^CKO, and PTEN^CKO + RG7388 mice under HCl+MV condition. Nuclei were counterstained with DAPI (blue). (K) Western blot analysis of E-cadherin, Vimentin and α-SMA protein expression in lung tissue from PTEN^F/F, PTEN^CKO, and PTEN^CKO + RG7388 mice under HCl+MV condition. (L-N) Quantification of E-cadherin, Vimentin and α-SMA expression levels normalized to GAPDH. Data are presented as mean ± SEM. n = 4 mice/group. (O) Immunofluorescence staining for E-cadherin (red), Vimentin (green), and α-SMA (green) in lung sections from PTEN^F/F, PTEN^CKO, and PTEN^CKO + RG7388 mice under HCl+MV condition. Nuclei were counterstained with DAPI (blue). One-way ANOVA and the Bonferroni post hoc test were used to compare more than two groups. **: P < 0.01, ***: P < 0.001, ****: P < 0.0001.