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. 2007 Jul 16;178(2):209–218. doi: 10.1083/jcb.200612031

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Copyright © 2007, The Rockefeller University Press

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Figure 3. — γ-H2AX is not removed from DNA by rapid turnover. (A) Cells were grown to log phase and treated with 0.1% MMS for 1 h. 10 or 20 mg/ml of caffeine was added at the same time when MMS was added to the culture. The activity of Mec1p and Tel1p kinases was examined by their ability to generate γ-H2AX, as shown by Western blot analysis. The CPY (carboxy peptidase Y) protein was used as a loading control. (B) A DSB was generated at MAT by HO induction. After 30 min, 10 mg/ml of caffeine was added into the cell culture to inhibit continued Mec1 and Tel1 kinase activity. γ-H2AX ChIP signals before HO induction as well as 0.5, 1, and 2 h after HO induction were examined.