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. 2021 Nov 19;49(21):12152–12166. doi: 10.1093/nar/gkab1068

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© The Author(s) 2021. 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 (https://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 6. — Relationship between histone H4K20 methylation levels and MCM hexamer states. (A) Representative microscopic images of Hoechst, H4K20me1, H4K20me2, and Mcm2. Scale bar, 20 μm. (B) Single-cell plot analysis of H4K20me1, H4K20me2, and Mcm2. The G1 cells with high levels of H4K20me1 (Hoechst < 1.125 and log₂(H4K20me1) > 0.3) are shown in light blue in the upper panel, those with low levels of H4K20me2 (Hoechst < 1.125 and log₂(H4K20me2) < −0.1) in blue in the middle panel, and those with high levels of Mcm2 (Hoechst < 1.125 and log₂(Mcm2) > 0.5) in pink in the lower panel. (C) Scatterplots between H4K20me1 and Mcm2 intensities, and between H4K20me2 and Mcm2 intensities based on (B). (D) Single-cell plot analysis of Mcm2, H4K20me2, and H4K20me3 in cells treated with either control-siRNA, or Suv420h1-siRNA. The number of cells examined in each panel is 450. (E) Western blot analysis using anti-Mcm2 (upper) and anti-Cdc45 (lower) antibodies following fractionation with a linear 5–30% sucrose gradient. The cells treated with Suv420h1-siRNA were used.