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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 7. — Xblm associates with replicating chromatin, but is not essential for DNA replication. (A) Sperm chromatin was incubated in egg extracts in the absence (lanes 1–5) or presence (lanes 6–10) of aphidicolin. At various times, chromatin fractions were isolated and immunoblotted for Xblm, RPA70, and Xorc2. (B) Xblm-depleted (lanes 1–4) and mock-depleted (lanes 5–8) egg extracts were incubated with sperm chromatin. Chromosomal DNA replication was assayed as described in the Materials and methods. The two bands in each lane depict ³²P incorporation into chromosomal DNA. (C) Xblm-depleted (lanes 1 and 2) and mock-depleted (lanes 3 and 4) extracts were incubated with sperm chromatin in the absence (lanes 1 and 3) or presence (lanes 2 and 4) of aphidicolin. Chromatin fractions were isolated and immunoblotted for Xblm, RPA70, and Xorc2. (D) Mock-depleted (lanes 1 and 2) and RPA70-depleted (lanes 3 and 4) extracts were incubated with sperm chromatin in the absence (lanes 1 and 3) or presence (lanes 2 and 4) of aphidicolin. Chromatin fractions were isolated and immunoblotted for Xblm, RPA70, and Xorc2. (E) Extracts were left untreated (lane 1) or were treated with either geminin (lane 2) or p27 (lane 3) as described previously (Lee et al., 2003), and sperm chromatin was added. After 90 min, chromatin fractions were immunoblotted for Xblm (top) and Xorc2 (bottom).