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. 2006 Apr;17(4):1897–1909. doi: 10.1091/mbc.E05-08-0727

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Copyright © 2006, The American Society for Cell Biology

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Figure 3. — Colocalization of VSV-tagged Survivin proteins and endogenous Aurora B in Survivin-depleted cells. U2OS cells were grown on glass coverslips and cotransfected with Survivin siRNA, plasmids expressing VSV-tagged versions of the indicated Survivin proteins and with H2B-GFP to visualize the DNA. Cells were fixed in paraformaldehyde 14 h after release from a thymidine block. (A) The exogenously expressed proteins were detected with mouse anti-VSV mAb (columns labeled VSV) and endogenous Aurora B with rabbit anti-Aurora B pAb (columns labeled Aurora B). The left panel shows prometaphases, and the right panel shows early and late anaphases. (B) Percentage of transfected (H2B-GFP pos.) cells with centromeric (red) or midbody (green) localization of the VSV-tagged Survivin proteins. For each mutant,50 prometaphases and 50 telophases were scored. In the Survivin_1-89 mutant, only 10 aberrant ana/telophases could be scored because the majority of these cells did not align their chromosomes nor underwent anaphase. (C) Percentage of transfected cells with centromeric localization of endogenous Aurora B. For each Survivin mutant, 75 prometaphases were scored.