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. 2014 Oct;24(10):1584–1594. doi: 10.1101/gr.173831.114

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© 2014 Ivanauskiene 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 5. — DEK depletion in ESCs induces fragile telomeres. Immunofluorescence detection of TERF1 (A) and telomere FISH analysis of telomere structure (B), on metaphase chromosomes from control and DEK-depleted cells. In control cells, TERF1 and telomere FISH show single signals of similar intensity on both sister chromatid ends. DEK depletion leads to chromatids with double or multiple TERF1 and telomere FISH signals with some stretched signals (arrows). (C) Proportions of metaphase spreads with fragile telomeres induced by DEK depletion (mean ± SD of three experiments; n = 105 cells). (D) Immunolocalization of gamma-H2AX in relation to telomeres (TERF1) in control and DEK-depleted cells. Arrows point to telomeres enriched in gamma-H2AX. Bars, 5 μm. (E) ChIP-qPCR analysis of gamma-H2AX enrichment at telomeres in control and DEK-depleted cells (mean fold enrichment of three experiments). Histones H3 and H4 were also chromatin immunoprecipitated as controls. (F) RT-qPCR analysis of telomere-encoded TERRA RNA transcripts in control and DEK-depleted cells.