Fig. 6

Bleomycin and TGF-β1 induced cell senescence in type II alveolar epithelial cells. (A, B) RLE-6TN cells were treated with a serial concentration of bleomycin, and Western blot analysis was performed to detect protein levels of p21 and p53. (A) representative blots. (B) Statistical analysis, n = 6. (C, D) After treatment with bleomycin for a serial of time points, Western blot was used to detect the protein expression of p53, p21, CDK6, and CDK2 in RLE-6TN. Statistical analysis was conducted. (E, F) Cells were treated with bleomycin (0.2 µg/ml) or TGF-β1 (2 ng/ml), and cell senescence levels were assessed by β-galactosidase assay. (G-J) Bleomycin or TGF-β1 was used to treat cells for 24 h, changes in cell cycle were detected by flow cytometry. (K, L) Senescence-related gene miRNA levels were tested using qRT-PCR. *P < 0.05, **P < 0.01, ***P < 0.001. P values were determined by the one-way ANOVA followed by the Bonferroni’s test