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. 2019 Nov 13;47(22):11637–11648. doi: 10.1093/nar/gkz1040

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© The Author(s) 2019. 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 5. — Thermal stability assay of the H3.1 Y41W–H4–DNA, H3.1 R63Q–H4–DNA and H3.1 F104M–H4–DNA complexes. (A) Reconstitution of the H3.1–H4–DNA, H3.1 Y41W–H4–DNA, H3.1 R63Q–H4–DNA and H3.1 F104M–H4–DNA complexes. H3.1, H3.1 Y41W, H3.1 R63Q and H3.1 F104M (lanes 2, 3, 4, and 5, respectively) were each mixed with the palindromic 145 bp satellite DNA fragment (lane 1), and the nucleosomes were reconstituted by the salt dialysis method. The reconstituted complexes were purified using a Prep Cell apparatus, and were analysed by 0.2× TBE nondenaturing 6% PAGE with ethidium bromide staining. (B) The histone contents of the purified H3.1–H4–DNA, H3.1 Y41W–H4–DNA, H3.1 R63Q–H4–DNA and H3.1 F104M–H4–DNA complexes were analysed by 16% SDS-PAGE with CBB staining (lanes 2, 3, 4, and 5 respectively). (C) The upper panel shows the normalised fluorescence intensity curves of the thermal dissociation of the H3.1–H4–DNA (•), H3.1 Y41W–H4–DNA (▓), H3.1 R63Q–H4–DNA (△) and H3.1 F104M–H4–DNA (◇) complexes. The bottom panel shows the derivative values of the thermal stability curves presented in the upper panel. The bars indicate standard deviations of triplicate experiments.