Anti phosphorylated p53

The tissues were homogenized in a lysis buffer. Fifty micrograms of proteins were loaded on a sodium dodecyl sulfate-polyacrylamide gel. The blots were probed with primary antibodies to polyclonal anti-hypoxic inducible factor-1 alpha (Hif-1a (diluted 1:1000; ab2185, Abcam, Cambridge, MA, USA), anti-phosphorylated p53 (diluted 1:500; #9284, Cell Signaling, Danvers, MA, USA), anti-Bax (diluted 1:500; #2772, Cell Signaling), and monoclonal anti-glucosetransporter (GLUT1; diluted 1:1000; ab40084, Abcam) at 4°C overnight. The primary antibody was visualized using secondary antibodies with an enhanced chemiluminescence kit (Amersham Pharmacia Biotech, Piscataway, NJ, USA).

A Cell Proliferation Kit I (MTT) was purchased from Roche Diagnostics GmbH (Mannheim, Germany). The antibodies used were anti-p38MAPK (14451, Cell Signal Technology, USA), antiphosphorylated p38MAPK (4511, Cell Signal Technology, USA), anti-MDM2 (86934, Cell Signal Technology, USA), antiphosphorylated MDM2 (3521, Cell Signal Technology, USA), anti-p53 (2527, Cell Signal Technology, USA), antiphosphorylated p53 (82530, Cell Signal Technology, USA), antiphosphorylated ATM (13050, Cell Signal Technology, USA), anti-ATM (2873, Cell Signal Technology, USA), antiphosphorylated H2AX (9718, Cell Signal Technology, USA), anti-H2AX (7631, Cell Signal Technology, USA), cleaved-caspase3 (9664, Cell Signal Technology, USA), and caspase3 (14220, Cell Signal Technology, USA). SB203580 and NSC207895 were purchased from Selleckchem (Houston, USA). β-Actin antisense oligonucleotides were purchased from Invitrogen (New York, USA). Human CGH4x72K Whole-Genome Tiling Array was purchase from Roche-NimbleGen (New York, USA). Proteome Profiler Human Phospho-MAPK Array Kit was purchased from R&D (Minneapolis, USA). Fluorescein isothiocyanate 4,6-diamidino-2-phenylindole (DAPI), diaminobenzidine (DAB) detection, and secondary antibody conjugated with horseradish peroxidase (HRP) were obtained from Zsbio (Beijing, China).