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. 2005 Apr;25(7):2660–2672. doi: 10.1128/MCB.25.7.2660-2672.2005

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Copyright © 2005, American Society for Microbiology

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FIG. 7. — E2F-induced senenescence is dependent on pRB, p16^Ink4a and p19^Arf. (A) E2F-induced senescence is pRB dependent. MEFs were passaged following a 3T3 protocol. Genotypes and treatments of cells were as follows: uninfected (RBloxP), ER-E2F3 infected (ER-E2F3, RB loxP), ER-E2F3 infected and induced with tamoxifen (ER-E2F3, RB loxP:OHT), Adeno-CRE infected (RB lox P: CRE), Adeno-CRE and ER-E2F3 infected and induced with tamoxifen (ER-E2F3, RB loxP: OHT and CRE). Tamoxifen administration and Adeno-CRE infection were performed at passage 5 and passage 6, respectively, as indicated by the arrows. (Inset) Protein expression levels of pRB in MEFs of the indicated genotype either infected with Adeno-CRE or not infected. The expression of vinculin was used as a loading control. (B) E2F3-induced senescence is partly dependent on p16^Ink4a. MEFswere passaged following a 3T3 protocol. Genotypes and treatments of cells were as follows: wild-type, ER-E2F3 infected (ER-E2F3, Ink4+/+), wild-type ER-E2F3 infected and induced with tamoxifen (ER-E2F3, Ink4+/+:OHT), p16^Ink4a null MEFs ER-E2F3 infected (ER-E2F3, Ink4*/*) p16^Ink4a null MEFs ER-E2F3 infected and induced with tamoxifen (ER-E2F3, Ink4*/*: OHT). Tamoxifen administration was performed at the indicated passage (arrow). (C) E2F-induced senescence is ARF dependent. MEFs were passaged following a 3T3 protocol. Genotypes and treatments of cells as follows: wild-type, ER-E2F3 infected (ER-E2F3, ARF+/+), wild-type ER-E2F3 infected and induced with tamoxifen (ER-E2F3, ARF+/+: OHT), ARF^−/− MEFs ER-E2F3 infected (ER-E2F3, ARF−/−), and ARF^−/− MEFs ER-E2F3 infected and induced with tamoxifen (ER-E2F3, ARF+/+: OHT). Tamoxifen administration was performed at the indicated passage (arrow).