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. 2017 Jun;27(6):913–921. doi: 10.1101/gr.215830.116

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© 2017 Delbarre et al.; Published by Cold Spring Harbor Laboratory Press

This article is distributed exclusively by Cold Spring Harbor Laboratory Press for the first six months after the full-issue publication date (see http://genome.cshlp.org/site/misc/terms.xhtml). After six months, it is available under a Creative Commons License (Attribution-NonCommercial 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/.

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Figure 1. — Loss of PML partitions histone H3.3 into a compact micrococcal nuclease (MNase)–resistant chromatin compartment. (A) MEF fractionation into a cytosolic fraction (S1), a Triton X-100 (0.5%)-insoluble/MNase (0.5 U/µg DNA)-soluble fraction (S2), and a Triton X-100/MNase-insoluble fraction (P2). (B) Nucleosome laddering (DNA; top) and Western blot (bottom) of endogenous H3.3 and total H3 in each fraction, from wt and Pml ko MEFs. (C) Proportions of H3.3 and total H3 in P2; (*) P < 0.05 relative to wt; paired t-tests. (D) Distribution of H3.3 and PML in S1, S2, and P2 from mock-transfected wt and Pml ko MEFs and from ko MEFs expressing MYC-PML1 or MYC-PML2 (full-length PML); alpha-tubulin was used as loading control for S1. (E) Proportions of H3.3 and total H3 in P2 after knockdown of Hira, Daxx, or Atrx; mean ± SD, three experiments; (*) P = 0.03 (siHira wt); 0.04 (siAtrx wt); 1.4 × 10⁻⁵ (siHira ko); (§) P = 0.06 (siDaxx, wt); paired t-tests relative to mock control.