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. 2009 May 22;284(21):14396–14404. doi: 10.1074/jbc.M808116200

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Copyright © 2009, The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 5. — ERK1/2 signaling mediates Bmi-1 down-regulation and chromatin dissociation in Atm^-/- astrocytes. A, Atm^+/+ astrocytes were synchronized by contact inhibition after subculture, at which time cells were treated with 100 μm H₂O₂. Bmi-1 association with chromatin was analyzed by fluorescence microscope for the presence of puncta in the nucleus. p16^Ink4a up-regulation and Bmi-1/chromatin dissociation was tested using anti-p16^Ink4a antibody. B, colocalization of Bmi-1 and 53BP1 was tested in the cells after H₂O₂ treatment using the same methods. C, Atm^+/+ astrocytes were left untreated or pretreated with 50 μm PD98059 (PD) overnight, before treating with 100 μm H₂O₂. The effects of 50 μm PD98059 on Bmi-1 localization in astrocytes were analyzed. D, photomicrographs of Atm^+/+ astrocytes culture from the experiment shown in C. Scale bars, 50 μm. E, Atm^-/- astrocytes were treated with 1 mm NAC or 50 μm PD98059 for 2 days. Bmi-1 expression and localization in Atm^+/+ and Atm^-/- astrocytes were determined. DAPI, 4′,6-diamidino-2-phenylindole.