Requirement of condensin in OA-dependent partial chromosome condensation in the interphase extract

Abstract

<p><b>Copyright information:</b></p><p>Taken from "Analysis of the role of Aurora B on the chromosomal targeting of condensin I"</p><p></p><p>Nucleic Acids Research 2007;35(7):2403-2412.</p><p>Published online 28 Mar 2007</p><p>PMCID:PMC1874644.</p><p>© 2007 The Author(s)</p> Mitotic (lanes 1–2) and interphase (lanes 3–4) extracts were mock-depleted (lanes 1 and 3) or depleted with a mixture of affinity-purified condensin antibodies (lanes 2 and 4). Equal volumes of these extracts were subjected to SDS-PAGE, blotted, and detected using the indicated antibodies. Sperm chromatin was incubated with mock-depleted mitotic extract (lanes 1–4), condensin-depleted mitotic extract (lane 5), mock-depleted interphase extract (lane 6), mock-depleted interphase extract supplemented with OA (lane 7), condensin-depleted interphase extract (lane 8) or condensin-depleted interphase extract supplemented with OA (lane 9) at 22°C for 2 h. The assembled structures were isolated, and 25 (lane 1), 12.5 (lane 2), 6.3 (lane 3), 3.1 (lane 4) and 100% (lanes 5–9) were analyzed by immunoblotting with the indicated antibodies. Sperm chromatin was incubated with mitotic extract (a), condensin-depleted mitotic extract (b), mock-depleted interphase extract (c), mock-depleted interphase extract with OA (d), condensin-depleted interphase extract (e), or condensin-depleted interphase extract with OA (f) at 22°C for 2 h. After incubation, assembled chromatin structures were fixed and stained with Hoechst. Bar, 10 μm