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. 2018 Aug 24;9:3420. doi: 10.1038/s41467-018-05912-z

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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. 5 — Knockdown of lamin B1 mainly affects non-NL integrated lamin B1. a qRT-PCR showing mRNA levels of lamin B1 in KD NMuMG cells. Expression levels were normalized to an endogenous control and expressed relative to the control-infected cells (C), which was set as 1. Data are presented as mean ± SD, n = 5. Left panel, representative western blotting for LB1 and tubulin (as a loading control) from C and KD NMuMG cells. Images are representative of three independent replicates (right). b Fractionation of nuclei from C and KD NMuMG cells to obtain cytoplasmic proteins, soluble proteins (NE1), and insoluble proteins (NE2), as monitored by western blotting using TBP as a NE1 control, tubulin as a cytoplasmic control, and H3 as a NE2 control. Images are representative of two independent replicates and five technical replicates. c Confocal fluorescence recovery after bleaching (FRAP) in C and KD cells expressing lamin B1-mCerulean. Circles denote the areas of bleaching. Each column displays prebleach, bleach, and recovery images. Rows 3 and 4 are magnifications of rows 1 and 2, respectively. Scale bar: 5 μm (inset 1 μm) (left). Images are representative of ten cells for each condition. Graphic representation of the relative intensity after 60 s of nuclear envelope and nucleoplasm recovery from bleaching in C and KD cells (right). Data are shown as mean, n = 6. d ChIP-qPCR of three selected lamin B1 + regions and a negative control (Oct4prom) selected from a negative lamin B1 + region, in C and KD NMuMG cells. Data from real-time PCR (qPCR) amplifications were normalized to the input and expressed as the fold-change relative to data obtained in C condition, which was set as 1. Data are shown as mean ± SD, n = 5. e ChIP-qPCR of three cLAD-selected regions in C and KD NMuMG cells. Data from qPCR amplifications were normalized to the input and expressed as the fold-change relative to data obtained in C condition, which was set as 1. Data presented as mean ± SD, n = 3

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