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. 2000 Jan 1;14(1):34–44.

Figure 3.

Figure 3

Effect of p53 on oxygen-regulated expression and stability of HIF-1α. (A) Immunoblot analysis of HIF-1α expression in p53+/+ and p53−/− HCT116 cells cultured for 8 hr in 20%or 1%O2. The blot was analyzed sequentially with monoclonal antibodies against HIF-1α (H1α67), p53 (DO-1), and β-actin. (B) Immunoblot analysis of HIF-1α expression in p53+/+ and p53−/− MEFs cultured for 8 hr in 20% or 1% O2. (C) Northern blot analysis of HIF-1α mRNA expression in p53+/+ and p53−/− HCT116 cells cultured as in A. (D). Immunoblot analysis of HIF-1α protein in p53−/− HCT116 cells cultured in 1% O2 for 8 hr following cotransfection with pCEP4–HIF-1α and either pCMV–p53 or empty vector. The blot was analyzed sequentially with anti-HIF-1α and anti-p53 monoclonal antibodies. (E) Half-life of HIF-1α protein in p53+/+ and p53−/− cells exposed to 100 μm cobalt chloride following addition of 100 μm cycloheximide. Lysates of cells harvested at the indicated time intervals were subject to immunoblot analysis of HIF-1α and p53 expression.