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. 2011 Apr 21;6(4):e19198. doi: 10.1371/journal.pone.0019198

Figure 4. NTERA-2D1 cells are hypersensitive to pro-apoptotic functions of p53 independent of DNA damage.

Figure 4

(A) Hypersensitivity of NTERA-2D1 cells is also observed upon DNA damage-independent accumulation of p53. Cells were treated with Cisplatin, Nutlin-3, or Bortezomib, respectively for indicated times and p53 protein was analyzed by Western Blot (left panel). In addition, cells treated for 24 h were stained with Annexin-V/FITC and PI and analyzed by flow cytometry (right panel). (B) Apoptosis upon the non-genotoxic agents Nutlin-3 and Bortezomib is dependent on p53. Cells were transfected with siRNA and cultivated for 48 h prior to Nutlin-3 or Bortezomib treatment (16 h). Cells were stained with Annexin-V/FITC and PI and analyzed by flow cytometry. Graph reflects means ±SD of survival after Cisplatin relating to corresponding controls from 3 experiments (P = 0.0003 and P = 0.0014 for Nutlin-3 and Bortezomib, respectively). (C) Nutlin-3 induced apoptosis directly correlates with the amount of p53 protein (determined by densitometry). Cells were transfected with indicated amounts of p53-targeting siRNA 48 h before Nutlin-3 treatment and analyzed by Western Blot.