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. 2005 Sep;25(17):7534–7545. doi: 10.1128/MCB.25.17.7534-7545.2005

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Copyright © 2005, American Society for Microbiology

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FIG. 2. — The histone chaperone activity of nucleophosmin is independent of its histone interaction. (A) Schematic depiction of the histone transfer assay to test the histone chaperone function of NPM1. (B) Histone transfer assay was carried out with 500 ng of histones and 250 ng of either full-length (FL) NPM1 or the deletion mutants. Lane 1, supercoiled DNA; lane 2, DNA relaxed by TopoI; lane 3, relaxed DNA with core histones; lane 4, relaxed DNA with core histones and full-length NPM1. Relaxed DNA without (lane 5) or with (lane 6) core histones and NPM1 1-200, relaxed DNA without (lane 7) or with (lane 8) core histones and NPM1 1-147, and relaxed DNA without (lane 9) or with (lane 10) core histones and NPM1 1-119 are shown. (C) Comparative analysis of the amount of supercoiled DNA and the histone H3-binding ability of NPM1 FL or each of the deletion mutants. The supercoiled DNA formed was quantified using Image Gauge software (Fuji) and expressed as a percentage of the total supercoiled DNA used. The percentage of histone H3 binding was calculated similarly.