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

Figure 6.

Figure 6

PTEN knockdown suppresses cell senescence in vitro and in vivo. (A) Schematic representation of the in vitro experimental procedure. BEAS-2B cells were transfected with or without PTEN siRNA and subjected to HCl + stretch. (B) Immunofluorescence staining for P53 (red) and P21 (red) in BEAS-2B cells under different conditions. Nuclei were counterstained with DAPI (blue). (C) Western blot analysis of P53 and P21 protein expression in BEAS-2B cells. (D-E) Quantification of P53 and P21 expression levels normalized to GAPDH. Data are presented as mean ± SEM. n = 4 /group. (F) Immunofluorescence staining for P53 (red) and P21 (red) in BEAS-2B cells following PTEN knockdown under HCl + Stretch condition. Nuclei were counterstained with DAPI (blue). (G) Western blot analysis of P53 and P21 protein expression in BEAS-2B cells following PTEN knockdown under HCl + Stretch condition. (H-I) Quantification of P53 and P21 expression levels normalized to GAPDH. Data are presented as mean ± SEM. n = 4 /group. (J) Schematic representation of the in vivo experimental procedure. Mice were subjected to HCl stimulation and mechanical ventilation (MV). (K) Immunofluorescence staining for P53 (red) and P21 (red) in lung tissue from mice under different conditions. Nuclei were counterstained with DAPI (blue). (L) Western blot analysis of P53 and P21 protein expression in lung tissue from mice under different conditions. (M-N) Quantification of P53 and P21 expression levels normalized to GAPDH. Data are presented as mean ± SEM. n = 4 mice/group. (O) Western blot analysis of P53 and P21 protein expression in lung tissue from PTENF/F and PTENCKO mice under HCl with or without MV conditions. (P-Q) Quantification of P53 and P21 expression levels normalized to GAPDH. Data are presented as mean ± SEM. n = 4 mice/group. (R) Quantification of IL-1β levels in bronchoalveolar lavage fluid (BALF) from mice under different conditions. Data are presented as mean ± SEM. n = 4 mice/group. (S) Quantification of IL-6 levels in BALF from mice under different 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.