Research Paper Volume 11, Issue 18 pp 7492—7509

PTEN loss regulates alveolar epithelial cell senescence in pulmonary fibrosis depending on Akt activation

Figure 7. Akt inactivation rescues AEC senescence in vitro. After PTEN was stably knocked down, an inhibitor of the AKT pathway (20 μm/L LY294002 or 5 μg/ml MK2206) was added 1 hour before bleomycin (10 μg/ml) to A549 cells for 72 hours followed by a fresh medium transfer for 24 hours. (A, B) Rescue of cell senescence by LY294002 was determined by SA-β-Gal staining (original magnification, 200×), and decreased expression of P21WAF1 (C, D) was confirmed by western blot. Rescue of cell senescence by MK2206 was determined by SA-β-Gal staining (E, F) and P21WAF1 expression (G, H). Data are shown as the mean ± SEM, n ≥ 3 per group. *p < 0.05, **p < 0.01. One-way ANOVA followed by Dunnett’s Multiple Comparison Test.