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Fig. 5. HP1{alpha} and aurora B kinase protein assemblies on HAC and extra-chromosomes, not at integrated sites in mitosis. (A-D) Metaphase chromosomes were immunofluorescent-stained with (A-D) specific antibodies and (B-D) by using a combination of FISH with the BAC probe. The localization of HP1{alpha} (red), compared with kinetochore protein CENP-C (green) in metaphase HT1080 cell is indicated in A. The presence or absence of HP1{alpha} (green in c-f, h, i) or aurora B kinase (green in g-j) on the BAC (red) locus was analyzed in (B) de novo HACs (S026), (C) the integration site (K031) or (D) the reformed minichromosome (K031) in metaphase-arrested cells. Chromosomes were counterstained with DAPI (blue or gray). Arrows indicate the HACs or the integration sites of the alphoid BAC. Arrowheads indicate canonical centromeres on the host chromosomes. Bars, 10 µm.





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