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Figure 8


Fig. 8. Mutation of Zeb1 does not trigger the classic Ink4A replicative senescence pathway in MEFs; instead, it is associated with induction of p15Ink4b and p21Cdkn1a. (A) Real-time PCR was used to compare mRNA expression from the Ink4a locus (p16Ink4a and Arf), and genes known to regulate the Ink4a locus (Ets1, Bmi1, Tbx2, Tbx3), in proliferating wild-type MEFs (P3) and senescent Zeb1 heterozygous (P5) and null (P2) cells. (B) p15Ink4b, p21Cdkn1a and E-cadherin mRNAs are induced in a gene dosage-dependent fashion in Zeb1 mutant MEFs. Real-time PCR results using the same samples as in A are shown. (C) ChIP assays. Input, starting chromatin used for the immunoprecipitations; IgG, preimmune serum. (D) Control ChIP assay showing that Zeb1 does not bind to the Gapdh promoter. Histone H3 and H4 are positive controls for binding to the Gapdh promoter. (E) Real-time PCR quantification of the results in C and D. The same input DNA was used for each ChIP assay, and the relative input value is set at 100. Primers for the promoters are shown in Table 2.





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