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. 2016 Jun 1;30(11):1313–1326. doi: 10.1101/gad.275073.115

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© 2016 Prendergast et al.; Published by Cold Spring Harbor Laboratory Press

This article, published in Genes & Development, is available under a Creative Commons License (Attribution 4.0 International), as described at http://creativecommons.org/licenses/by/4.0/.

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Figure 3. — The Spt16-CTD is sufficient to bind CENP-T/-W in vitro. (A) Scheme depicting domains of Spt16 (amino acids within the respective domains summarized in Winkler and Luger 2011). The regions of Spt16 that were expressed as GST-tagged recombinant proteins are illustrated. Recombinant GST-tagged Spt16 fragments were used as bait in a GST pull-down assay. CENP-T^HFD/-W was used as prey. GST alone was used as a control for pull-downs. Mutants that contained Spt16 amino acids 926–965 could bind CENP-T^HFD/-W. (B) Schematic illustrating the amino acids within Spt16 (amino acids 926–965), which were mutated to alanine and expressed as recombinant GST-tagged proteins. The Coomassie gel shows input samples and GST pull-downs using GST-tagged Spt16 mutants as bait, and CENP-T^HFD/-W was used as prey. GST alone was used as a pull-down control. The bar chart shows quantification of the amount of CENP-T/-W pulled down by each Spt16 mutant in two independent experiments.