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. 2019 Feb 4;38(22):4232–4249. doi: 10.1038/s41388-019-0723-8

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© The Author(s) 2019

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. 8 — Cyclin D1 determines accumulation of G9a-mediated nuclear lamina interactions at the nuclear periphery. a Illustration of the inducible ^m6A-tracer/Dam-lamin B1 system in the HT1080 cell line stably expressing the Shield1-inducible Dam-Lamin B1 and the Tet-off ^m6A -tracer (known as the line clone 3). Inducible Dam-Lamin B1 expression was established by the fusion of a destabilization domain (DD), which causes Dam-Lamin B1 to be rapidly targeted for proteasomal degradation unless the protein is shielded by the synthetic small molecule Shield1.The induction of ^m6A-tracer is based on the Tet-Off system, where the removal of doxycycline (Dox) results in the activation of transcription. b Schematic representation of the experimental protocol. c Confocal microscopy images showing both G9a shRNA/GLP shRNA and cyclin D1 siRNA decreases H3K9me2 staining (red) in H1080 cells. d Representative confocal microscopy images showing cyclin D1 siRNA decreases accumulation of ^m6A-tracer signal (green) at the nuclear periphery after induction of Dam-Lamin B1. Lamin B1 staining is shown in red. e Quantitative analysis of ^m6A-tracer incorporation at the nuclear periphery after induction of Dam-Lamin B1 expression shown as % of cells incorporating tracer for n > 200 separate cells for shG9a/shGLP treatment and its vector control. Data are percentage ± 95% confidence interval (CI). f Quantitative analysis of ^m6A-tracer incorporation at the nuclear periphery shown as % of cells incorporating tracer for n > 400 separate cells cyclin D1 siRNA treatment as well as its control. Data are percentage ± 95% CI