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. 2004 Jun 21;165(6):801–812. doi: 10.1083/jcb.200402095

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Copyright © 2004, The Rockefeller University Press

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Figure 1. — Xblm binds to chromatin and becomes phosphorylated in a checkpoint-dependent manner. (A) Extracts were incubated with no sperm chromatin (lane 1), untreated sperm chromatin (lane 2), sperm chromatin plus 100 μg/ml aphidicolin (APH; lane 3), UV-damaged sperm chromatin (lane 4), or sperm chromatin plus aphidicolin and caffeine (lane 5). After 90 min, nuclei were isolated and separated into nuclear soluble and chromatin fractions. The fractions were immunoblotted for Xblm and Xorc2 (as a loading control). (B) Extracts were incubated for 90 min with untreated sperm chromatin (lane 1), UV-damaged sperm chromatin (lane 2), or sperm chromatin plus aphidicolin (lane 3). Whole nuclear fractions were immunoblotted for Xblm. (C) Extracts were incubated with sperm chromatin alone (lane 1), sperm chromatin plus aphidicolin (lane 2), or sperm chromatin plus aphidicolin and caffeine (lane 3). Nuclear fractions were immunoblotted for Xblm. (D) Anti-Xblm immunoprecipitates from extracts containing no DNA (lane 1) or pA-pT (lanes 2 and 3) were incubated without (lanes 1 and 2) or with (lane 3) λ phosphatase and were immunoblotted for Xblm.