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Figure 2


Fig. 2. Condensin I binding to chromatin is dependent on Aurora B activity. (A) Nocodazole-arrested human HeLa cells were either treated with 100 nM Hesperadin or DMSO for 1 hour, fixed and stained for the condensin subunits Smc2 (shared between condensin I and II), kleisin-{gamma}, Cap-D2 (both condensin I specific) and Cap-D3 (condensin II specific). Chromosomes were counterstained with DAPI (1 µg/ml). Bar, 10 µm. (B) Average fluorescence intensities of condensin on chromatin were quantified using the DAPI channel as a reference and to determine the area occupied by chromatin. The background intensity was calculated by measuring the fluorescence intensity outside this area and subtracted from the condensin signal. The presented graphs are averaged from at least 30 different cells for each condition (n>30) and normalised to the control value ± s.d. (C) Biochemical fractionation of Nocodazole-arrested HeLa cells which had either been treated with 100 nM Hesperadin for 1 hour or left untreated before harvesting. The levels of condensin-I-(Cap-G and kleisin-{gamma}) and condensin-II-specific subunits (Cap-D3 and Cap-G2) were detected by western blotting. Total mitotic extracts (ME), cytoplasm (Cyt) and chromatin (Chr) fractions are shown.





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