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. 2023 Jan 20;51(4):1674–1686. doi: 10.1093/nar/gkac1274

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© The Author(s) 2023. 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-NonCommercial License (https://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com

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Figure 7. — Opposite effects of mutagenesis on DNA binding by removing or introducing acidic residues within the hairpin loop. (A) The isolated NT fragment (residues 1–216) does not bind to DNA but the NT-ZF fragment (residues 1–366) has reduced DNA binding potential (left) (N = 7). An 18% SDS-PAGE stain-free gel (right panel) showing the recombinant protein samples used in DNA binding assays. (B) The isolated NT fragment added in trans does not prevent the DNA from binding to the ZF array. (C) The mutant with four D/E-to-A substitutions (red line) within the hairpin loop increased DNA binding in the context of full-length ZNF410. (D) The mutant with seven S/T-to-D/E substitutions (black line) within the hairpin loop further reduced DNA binding in the context of full-length ZNF410 (N = 6). ZF, blue lines and FL, brown lines.