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. 2011 Dec;31(24):4928–4937. doi: 10.1128/MCB.06085-11

Fig. 1.

Fig. 1.

Effect of hypoxia on MDM2 modification and induction of VEGF. (A) Expression of MDM2, p53, and VEGF proteins in four cultured neuroblastoma cell lines indicated above the blots, plus the pair of MDM2+/+ p53−/− and MDM2−/− p53−/− MEF cells, all detected by Western blotting. The positions of molecular mass markers (in kilodaltons [kd]) are indicated to the left of the blots. (B) SHEP cells exposed to hypoxia for different times (in hours) as indicated above the top blot. Cell extracts were tested by Western blot assay for MDM2, phosphorylated MDM2 (p166), and VEGF expression in whole-cell extract (WCE), as well as for MDM2 in nucleus (Nuc) and cytoplasm (Cyt) fractions. The values under protein bands in the blots represent expression levels after normalization for controls (GAPDH or nucleolin and tubulin), compared with untreated (0) samples (defined as 1 unit). (C and D) Comparison of VEGF mRNA induction during hypoxia in MDM2-positive versus MDM2-negative cells. SHEP (MDM2-positive) and SK-N-SH (MDM2-negative) cells that have wt p53 (C), and both MDM2+/+ p53−/− and MDM2−/− p53−/− MEF cells (D) were exposed to hypoxia for different times, as indicated, and VEGF mRNA expression was detected by qRT-PCR.