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Fig. 6. Western blot analysis demonstrates that elevated expression of STAT-1 is associated with enhanced expression of MDC1 and 53BP1, and constitutive phosphorylation of ATM, Chk2 and NSB1 in cells that lack p53 or carry a mutation for p53. (A) Lysates of Soas2 (p53-deficient), HCT15 (p53 mutant), IMR90 (p53 wild type) or SKNSH (p53 wild type) cells were immunoblotted with antibodies against the proteins indicated. (B) STAT-1 levels are enhanced in MEF p53^–/– cells compared with MEF p53^+/+ cells as assessed in a western blot. (C,D,E) Western blots show that, transfection of HCT15 and Soas2 cells (C and D, respectively) with STAT-1 RNAi reduces levels of STAT-1 protein, levels of MDC1 and 53BP1, and also reduces the level of phosphorylated ATM (pATM-S1981). Overexpression of STAT-1, by contrast, increases levels of phosphorylated ATM and the phosphorylated forms of downstream phosphorylation-substrates of ATM. IMR90 cells were transfected with a STAT-1 expression vector; cells were harvested 48 hours later and lysates immunoblotted with antibodies against the proteins indicated.

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