Research Paper Volume 12, Issue 18 pp 18501—18521

D-galactose induces senescence of glioblastoma cells through YAP-CDK6 pathway

Figure 7. Overexpression of YAP restored D-gal-induced GBM senescence. (A) A schematic illustration showing YAP transfection in C6 senescent cells. (B) Immunostaining of PH3 in control C6 cells without transfection, or senescent C6 cells (treated with 222 mM D-gal for 8 d) transfected with EGFP or YAP-EGFP plasmid (YAP-Res) for 2 d. (C) Quantitative analysis of the percentage of PH3+ cells over total C6 cells as shown in (B) (n=15). (D) Representative images showing β-galactosidase staining in control C6 cells without transfection, or senescent C6 cells (treated with 222 mM D-gal for 8 d) transfected with EGFP or YAP-EGFP plasmid (YAP-Res) for 2 d. (E) Quantification of the percentage of β-galactosidase+ cells over total cells as shown in (D) (n=15). (F) Western blot detected the expression of YAP, CDK6, and Lamin B1 in control C6 cells without transfection, or senescent C6 cells (treated with 222 mM D-gal for 8 d) transfected with EGFP or YAP-EGFP plasmid (YAP-Res) for 2 d. (G, H) Quantification of CDK6 and Lamin B1 level as shown in (F) (n=10). Scale bars, 20 μm. Data shown are mean ± s.e.m. *P < 0.05, **P < 0.01. ***P < 0.001.