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. 2013 Apr 24;41(11):5769–5783. doi: 10.1093/nar/gkt314

Figure 5.

Figure 5.

Cse4 hemisomes are stable in high concentrations of urea. (A and B) Native 6% gelFRET showing gel-shifted particles after dialysis versus 10 mM HEPES (pH7.5) + 0.25 mM EDTA containing the indicated concentrations of urea, using H3 octamers (A) and Cse4 octamers (B). The slowly migrating Cse4 band varies between experiments (see Supplementary Figure S7) and might represent an intact pseudo-octasome, a ‘stack’ of hemisomes or a stable aggregate. (C) Cse4 octamers and α62 duplexes were dialyzed versus 4 M urea + 10 mM HEPES (pH 7.5) + 0.25 mM EDTA using different combinations of end-labeled fluorophores as indicated, where Cy or Al on left indicates a 5′ label on α62 F oligonucleotide, and on right for α62 R. For each lane, a DNA band corresponding to the band marked by asterisks in Figure 3A is shown in the middle image and served as a negative control. The lower image shows gel shifts for labeled α62 duplexes that had been diluted 5-fold with unlabeled α62 duplexes.