Fig. 2

MLN8237 stimulated cellular senescence was mediated by P53 pathway. Senescence-associated signal pathways were detected by western blot. IMR32 cells were treated with 2 μmol/l of MLN8237. DMSO or no treatment as control. Cell samples were collected at 6 h, 12 h, and 24 h. Total proteins were extracted for western blot analysis for (a) p16, Rb, pRb (807), and pRb (796); b P27, PTEN, p53, and p21. c IMR32 cells were treated with 0, 0.5, and 2 μmol/l of MLN8237, respectively. At 6 h and 24 h, cells were collected, and total proteins were extracted for western blot analysis for p53 and p21. d At 48 h after MLN8237 treatment, IMR32 cells were collected and total RNA were extracted. P21 mRNA were quantified by real-time RT-PCR