Fig. 1.

EMS is negatively regulated by p53. (A) Real-time RT-PCR analysis of EMS levels in A549 and H460 cells infected with lentiviruses expressing control shRNA or p53 shRNA. Data shown are mean ± SD (n = 3). LE and SE indicate long-time exposure and short-time exposure, respectively. *P < 0.05; ***P < 0.001. (B) Real-time RT-PCR analysis of EMS levels in A549 and H460 cells infected with lentiviruses expressing control (pCDH) or p53 (pCDH–p53). pCDH is a lentiviral expression vector. Data shown are mean ± SD (n = 3). *P < 0.05. (C–E) A549 cells expressing control shRNA or p53 shRNA were treated with (C) 10 μM Etoposide, (D) 0.5 μg/mL Doxorubicin, or (E) 10 μM Nutlin-3 for 24 h. Total RNA and cell lysates were then analyzed by real-time RT-PCR and western blotting, respectively. Data shown are mean ± SD (n = 3). *P < 0.05; ***P < 0.001. (F) EMS was expressed at relatively lower levels in LUAD with wild-type (WT) p53 (n = 259) than in those carrying mutated (MUT) p53 (n = 248). (G) A549 cells were treated with pifithrin-α (PFT-α) (20 μM) for the indicated periods of time, followed by real-time RT-PCR and western blot analyses. Data shown are mean ± SD (n = 3). *P < 0.05; **P < 0.01. (H) H1299 cells were transfected with plasmids encoding wild-type p53 or the indicated mutant forms of p53. Twenty-four hours after transfection, total RNA and cell lysates were analyzed by real-time RT-PCR and western blotting, respectively. Data shown are mean ± SD (n = 3). ns., no significance. *P < 0.05.