Research Paper Volume 12, Issue 18 pp 18501—18521

Figure 9. Metformin reversed D-gal-induced senescence of GBM cells by activating YAP-CDK6 pathway. (A) A schematic illustration showing metformin treatment in the presence of D-gal. (B) Immunostaining of PH3 in control C6 cells, or senescent C6 cells (treated with 222 mM D-gal for 8 d), or senescent C6 cells (treated with 222 mM D-gal for 8 d) with metformin treatment (2 mM for 1 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, senescent C6 cells (treated with 222 mM D-gal for 8 d), or senescent C6 cells (treated with 222 mM D-gal for 8 d) with metformin treatment (2 mM for 1 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, or senescent C6 cells (treated with 222 mM D-gal for 8 d), or senescent C6 cells (treated with 222 mM D-gal for 8 d) with metformin treatment (2 mM for 1 d). (G–I) Quantification of YAP (n=6), CDK6 (n=15), and Lamin B1 (n=8) level as shown in (F). Met: metformin. Scale bars, 20 μm. Data shown are mean ± s.e.m. ^*P < 0.05, ^**P < 0.01, ^***P < 0.001.