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A single Cse4 nucleosome forms over the CEN. MNase-treated chromatin prepared from strain SBY5146 (FLAG-CSE4) was immunoprecipitated with anti-FLAG agarose beads. Cse4-associated DNA was isolated and separated on a 1.5% agarose gel. Southern blotting was performed with probes specific to either the CEN3 or a CEN3-flanking nucleosome. M, radiolabeled 100-bp ladder marker; IP, the DNA bound to Cse4.
