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Figure 2


Fig. 2. R246S mutant protein exhibits DN effects in ES cells. (A) The expression of p53 target genes noxa, p21 and mdm2 was analyzed by quantitative real-time PCR 6 hours after doxorubicin (0.5 µg/ml) treatment. The expression level of each gene was normalized with the expression of gapdh and fold induction was calculated. Data represents mean ± s.e.m. of four independent experiments, each performed in duplicate. (B) Expression of mdm2 mRNA was analyzed by northern blot hybridization after doxorubicin (0.5 µg/ml) or {gamma}-irradiation (5 Gy). Intensity of 28S ribosomal RNA on agarose gel is shown for the loading control. (C,D) Cell-death analysis upon genotoxic stresses. Undifferentiated ES cells were treated with various doses of doxorubicin (C) or UV (D) for 12 hours and cell death was determined by flow cytometry after staining with annexin-V/propidium iodide. Data represents the percentage survival normalized to untreated samples (mean ± s.e.m.) from four independent experiments, each performed in duplicate.





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