Research Paper Volume 12, Issue 18 pp 18501—18521

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

Figure 8. Overexpression of CDK6 restored D-gal-induced GBM senescence. (A) Immunostaining of PH3 in control C6 cells without transfection, or senescent C6 cells (treated with 222 mM D-gal for 8 d) transfected with pcDNA3.1 or pcDNA3.1-CDK6 plasmid (CDK6-Res) for 2 d. (B) Quantitative analysis of the percentage of PH3+ cells over total C6 cells as shown in (A) (n=15). (C) 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 pcDNA3.1 or pcDNA3.1-CDK6 plasmid (CDK6-Res) for 2 d. (D) Quantification of the percentage of β-galactosidase+ cells over total cells as shown in (C) (n=15). (E) Western blot detected the expression of 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 pcDNA3.1 or pcDNA3.1-CDK6 plasmid (CDK6-Res) for 2 d. (F) Quantification of Lamin B1 level as shown in (E) (n=10). Scale bars, 20 μm. Data shown are mean ± s.e.m. *P < 0.05, ***P < 0.001.