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. 2008 Dec 22;205(13):3007–3018. doi: 10.1084/jem.20081165

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© 2008 Urbonaviciute et al.

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.jem.org/misc/terms.shtml). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/).

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Figure 3. — HMGB1 cofractionates with nucleosomes from apoptotic cells. For induction of apoptosis, Jurkat cells were treated with 1 μM staurosporine for 8 h. Necrosis was induced by heat treatment at 56°C for 30 min. Nuclei isolated from viable (A), necrotic (B), and apoptotic (C) Jurkat cells were digested with micrococcal nuclease, lysed, and fractionated by sucrose gradient ultracentrifugation. Eight fractions were collected in each gradient from top (1) to bottom (8), and individual fractions were analyzed by 1.5% agarose gel electrophoresis in the presence of 1% SDS. HMGB1 was detected by Western blotting. (D) Characterization of mononucleosomes. 10 μg of total protein of mononucleosomal fraction 4 purified from necrotic (lane 1), viable (lane 2), and apoptotic (lane 3) Jurkat cells was subjected to a 15% SDS-PAGE. The proteins were visualized by Coomassie blue staining.