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. 2001 Jan 23;98(3):1036–1041. doi: 10.1073/pnas.021282898

Figure 3.

Figure 3

DNA replication block actively represses the p53 activating pathway. (A) RKO, HCT116, GM02186, and WI38 cells were not treated (−) or exposed to HU treatment for 24 h (HU and HU + γ) or subjected to γ IR for 12 h (HU + γ and γ) before lysis, and subjected to Western blot analysis using p53, p21, and actin probes. (B) RKO cells were treated as in A or exposed to APH treatment (5 μg/ml) for 24 h (Aph and Aph + γ) or subjected to γ IR (Aph+ γ and γ) for 12 h before lysis. Extracts were subjected to Western blot analysis of p53 protein using Pab 1801(p53) and modification-specific antibodies as described in the text. (C) The same extracts as in B were used for Western blot analysis of the p53 downstream targets: p21, HDM-2, and PIG3. Actin was used as a loading control.