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Figure 5


Fig. 5. Immunofluorescence of topo-II{alpha}-expressing and topo-II{alpha}-depleted (72 hours exposure to dox) HTETOP cells. (A) Metaphases (cytospun, no colcemid) stained using an antibody against CENP-A (green); (B) partial chromosome spreads (colcemid) stained for CENP-F (green); (C) partial metaphase spreads (colcemid) stained for CENP-E (green); (D) typical metaphase cells (grown in situ, no colcemid) stained for CENP-C (green) and Aurora B (red); (E) immuno-FISH of partial chromosome spreads (colcemid) showing the typical banding pattern generated using an antibody raised against acetyl-histone H3 (amino acids 1-20) (green). The centromeric regions have been identified using a pan-alphoid DNA probe (red). The white arrowheads highlight examples of hypoacetylated centromeric and pericentromeric domains. DNA is counterstained with DAPI (blue). Bars, 10 µm.