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. 2011 Jul 26;30(16):3397–3415. doi: 10.1038/emboj.2011.248

Figure 3.

Figure 3

The full-length and DNA-binding site of p53 were required for p53 suppression of BNIP3. (A) The schematic depiction of the p53 mutants. (B) The expression of the p53 mutants was verified by western blot using a monoclonal antibody against HA, GAPDH was used as a loading control. (C) The suppressive activity of p53 mutants on the BNIP3 promoter was measured by promoter assays under normoxia in Hela cells. (D) The suppressive activity of p53 mutants on the BNIP3 promoter was evaluated by promoter assays under hypoxia in Hela cells. (E) The expression of HA-tagged wild-type p53 and DNA-binding site mutated p53 (R175) was verified by western blot using anti-HA antibody under normoxia and hypoxia. (F) The suppressive activity of wild-type p53 and mutated p53 (R175H) on the BNIP3 promoter was checked by promoter assays under normoxia. (G) The suppressive activity of wild-type p53 and mutated p53 (R175H) on BNIP3 promoter was determined by promoter assays under hypoxia. Data of luciferase reporter assays are reported as mean±s.d. of three independent experiments performed in triplicate; the statistical analysis was performed using GraphPad Prism 5.0 (t-tests).