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. Author manuscript; available in PMC: 2011 May 11.
Published in final edited form as: Methods. 2006 Dec;40(4):312–317. doi: 10.1016/j.ymeth.2006.06.023

Figure 2.

Figure 2

Assays for nucleosome sliding activity of the human INO80 complex. Human INO80 complex was prepared as described in the text from cells expressing FLAG-hIes2 (PAPA-1). Panel A. Reactions contained ~500 ng INO80 complex, 10 fmol mononucleosomes reconstituted on the 216 bp dSH-A fragment, which does not contain a nucleosome positioning sequence, and ~270 fmol of long oligonucleosomes, and either 1 mM ATP or 1 mM ATPγS. The starting mononucleosomes are a mixture of laterally positioned (lower band) and centrally positioned (upper band) mononucleosomes. At the conclusion of the reactions shown in lanes 1-3, a mixture of long oligonucleosomes and salmon sperm DNA was added to compete bound INO80 complex off the labeled mononucleosomes and free DNA. Panel B. Reactions contained ~500 ng INO80 complex, 10 fmol mononucleosomes reconstituted on a 212 bp DNA fragment with a 601.2 nucleosome positioning sequence at one end, and the indicated amounts of short oligonucleosome competitor, prepared as described [15].