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. 2012 Jul 15;26(14):1533–1545. doi: 10.1101/gad.184911.111

Figure 2.

Figure 2.

E2F7 is up-regulated in a p53-dependent manner in response to either Nutlin3a treatment or ectopic expression of p53. (A) Immunoblot analysis of E2F7, p53, p21, and Actin expression using cell extracts from U2OS cells nontreated and treated with increasing amounts of the Mdm2 inhibitor Nutlin3a. (B) Cell cycle profile using flow cytometry of corresponding samples in A. (C) qRT–PCR expression analysis of TP53, CDKN1A, E2F6, E2F7, and E2F8 using mRNA extracted from U2OS p53+/+ cells nontreated or treated with increasing amounts of Nutlin3a. (D) Immunoblot analysis of p53 levels using the EJ cell line, which was engineered to ectopically express p53 under the control of a tetracycline-repressible system. (E) Cell cycle profile analysis of each corresponding sample in D before and after removal of tetracycline from cells in culture. (F) qRT–PCR expression analysis of the same target genes used in C from each corresponding EJp53 sample shown in D and E. The average of three independent experiments is shown. (NT) Nontreated; (T) tetracycline. Error bars indicate the standard error of the mean. Statistical significance is shown using the Student's t-test analysis; (**) P < 0.01; (*) P < 0.05; (n.s.) not significant.