摘要
Figure 4.AKT is phosphorylated in p53>dependent manner in ALT cells.(A)Western blot determination of total and phosphorylated AKT(S473)in p53-positive(VA13,U20S)and p53-defective(SAOS2,SKLU-1)ALT cells.(B)Knockdown of p53 in U20S or moderate expression of p53 in SAOS2 induces down or up-regulation of p-AKT,respectively.(C)Knockdown of ATM or ATR by siRNA decreases abundance of p53,phosphorylated p53 and p-AKT.(D)Quantitative-PCR determination of the level of ATR or ATM in U20S cells transfected with siRNA to ATR or ATM,respectively.Scramble siRNA(Si-Ctl)was used as control.Data represent the mean±SEM,n=3-4.(E)ATM(KU60019)or ATR(VE-821)inhibitor decreases abundance of p-AKT in U20S cells.U20S cells were treated with indicated concentration of KU60019 or VE-821 for 24 h.(F)The expression of wt-p53,but not mutant p53(p53-s269e)defective of transcription activity,increases the level of p-AKT.(G)PFTa.an inhibitor of p53 transcription activity,suppresses the phosphorylation of AKT.U20S cells were treated with indicated concentration of PFTa for 24 h.
