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. 2015 Apr 27;43(10):4909–4922. doi: 10.1093/nar/gkv405

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© The Author(s) 2015. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 4. — The CENP-B DBD specifically binds to the CENP-A-H4 complex. (A) Schematic representation of the pull-down assay with the His₆-tagged CENP-B DBD and the CENP-A-H4 or H3.1-H4 complex. (B) The His₆-tagged CENP-B DBD (50 nM) complexed with a 21 base-pair DNA was incubated with the CENP-A-H4 or H3.1-H4 complex (50 nM). The proteins pulled down with the Ni-NTA agarose beads were analyzed by 16% SDS-PAGE with Coomassie Brilliant Blue staining. Lane 1 indicates the molecular mass markers. Lane 2: His₆-tagged CENP-B DBD (50% of input). Lane 3: the H3.1-H4 complex (20% of input). Lane 4: the CENP-A-H4 complex (20% of input). Lanes 5 and 6 represent the pull-down experiments with the H3.1-H4 complex, in the absence and presence of the His₆-tagged CENP-B DBD, respectively. Lanes 7 and 8 represent the pull-down experiments with the CENP-A-H4 complex, in the absence and presence of the His₆-tagged CENP-B DBD, respectively.