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. 2006 Dec 18;175(6):869–880. doi: 10.1083/jcb.200604005

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Copyright © 2006, The Rockefeller University Press

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Figure 5. — Histone H1 is lost from various types of senescent cells. (A) Mock- or RasG12V-transfected cells (WI-38, MRC-5, IMR-90, and BJ) were fractionated as shown in Fig. 4 A at day 5 after drug selection. AE and cytoplasmic fractions (C) were analyzed by immunoblotting. The amounts of loaded proteins were calibrated by anti-H2B or antiactin signals, respectively. Transfected cells were scored for the percentages of BrdU- and SAHF-positive cells at harvest time (bottom; n = 200). (B) WI-38 cells were induced to senesce by the retroviral expression of RasG12V or a constitutive active form of MKK6 (MKK6EE). Replicative senescent cells were analyzed at 57 population doubling levels, at which cells could not reach confluence for 3 wk. Quiescent cells were prepared by culturing the mock-transfected cells in 0.1% FBS for 1 wk. WCEs were analyzed by immunoblotting. The percentages of BrdU- and SAHF-positive cells are indicated (bottom; n = 200).