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Fig. 8. Examination of CAP-D2 and DNA topoisomerase II in metaphase chromosomes after dsRNA treatment. (A,B) Metaphase chromosome spreads of control (1,2), topo II-depleted (3), CAP-D2-depleted (4) and CAP-D2/Topo II doubly depleted (5) cells, after cells were arrested with colchicine. Cells were cytospun onto poly-L-lysine slides, extracted during fixation and stained for topo II (red), CAP-D2 (green) and DNA (blue), 74 hours after dsRNA treatment. (A) Control cells displayed the typical X-shape and sister chromatids could be easily distinguished. Both CAP-D2 and topo II localised axially along the length of the chromosomes with partial colocalisation (A1,2 and B, Control). In topo II, CAP-D2 single depletions, and in the CAP-D2/Topo II double depletion, no sister chromatid resolution could be observed and chromosomes appeared fuzzy. Scale bar: 10 µm. (B) Individual chromosomes observed in chromosome spreads. The chromosome phenotype was similar in the two single and the double RNAi experiments.





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