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. 2018 Dec 3;9:5143. doi: 10.1038/s41467-018-07189-8

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© The Author(s) 2018

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 2 — MacroH2A1 localization to H2Bac chromatin is required for regulation of H2BK12ac, H2BK120ac, and SASP gene transcription. a Diagram illustrates macroH2A1.1-dependent positive feedback loop regulating the induction of SASP gene expression. Senescent stimuli, such as H-Ras^V12 upregulates the expression of macroH2A1.1 which promotes the transcription of SASP genes. Secreted SASP factors can act in an autocrine or paracrine fashion. Conditioned media from senescent cells contains secreted cytokines and chemokines (SASP factors) which can promote senescence through paracrine signaling. b Schematic of domain swap macroH2A1.1-Flag (mH2A) mutant lentiviral constructs expressed in IMR90-hTERT cells. MacroH2A1.1 exon and docking domain indicated. c Immunoblots of acid-extracted histones for indicated histones and histone PTMs from cells that ectopically express macroH2A1.1 constructs described in b. d RT-qPCR for SASP factors of cells described in b. Error bars, s.e.m. (n = 3 independent lentiviral infections, *p < 0.05 from a two-tailed Student’s t-test). e Flag ChIP-qPCR in macroH2A1.2-Flag cells for SASP genes shown in d. Bars represent enrichment of macroH2A1-Flag at SASP loci over unbound control regions. Error bars, s.e.m. (n = 3 independent cell passages, *p < 0.05 from a two-tailed Student’s t-test)