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. 2017 Jun 12;8:15616. doi: 10.1038/ncomms15616

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Copyright © 2017, The Author(s)

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(a) Experimental scheme to determine the efficiency of INO80 for dimer exchange at single-nucleosome level, based on three-colour FRET. Nucleosomes forming FRET pairs with Cy3 (DNA) and Cy5.5 (H2A–H2B dimer) or ATTO 647N (H2A.Z–H2B dimer) were counted to measure the efficiency of dimer exchange. The acceptor is photo-bleached at a single time point (black arrow), which is followed by a single-step donor (Cy3) photobleaching (red arrow). (b,c) The results are shown for efficiencies of H2A–H2B incorporation and H2A.Z–H2B incorporation (b), and the effects of a single nucleotide gap or nick at nt −58 or the absence of ATP on H2A–H2B incorporation (c). The number of single molecules (n) analysed for each reaction condition is shown above its corresponding bar.