Figure 3. D-gal induced changes of senescence makers in C6 cells. (A) Western blot detected the expression of p53 and Lamin B1 in C6 cells treated with 0 mM, 28 mM, 111 mM, and 222 mM D-gal for 8 d. (B, C) Quantification of p53 (n=7) and Lamin B1 (n=5) expression as shown in (A). (D) Immunostaining analysis of Lamin B1 (green) in control and senescent C6 cells (treated with 222 mM D-gal for 8 d). (E) qPCR analysis of p16 mRNA level in control cells and senescent C6 cells (treated with 222 mM D-gal for 8 d) (n=3). (F) qPCR analysis of NF-κB mRNA level in control cells and senescent C6 cells (treated with 222 mM D-gal for 8 d) (n=3). (G) Western blot detected the expression of NF-κB and p53 in control cells and senescent C6 cells (treated with 222 mM D-gal for 8 d). (H) Quantification of NF-κB expression as shown in (G) (n=12). Scale bars, 20 μm. Data shown are mean ± s.e.m. *P < 0.05, **P < 0.01, ***P < 0.001.