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. 2017 Feb 23;45(8):4281–4293. doi: 10.1093/nar/gkx125

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© The Author(s) 2017. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com

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Figure 4. — Overexpression of POT1 partially rescues the telomeric DNA damage response. CTC1 conditional cells that did/did not overexpress exogenous POT1-FLAG were monitored for POT1-FLAG, γH2AX and RPA localization to telomeres after growth for 7 days with/without tamoxifen. (A) Localization of POT1-FLAG to telomeres. Interphase cells were analyzed by telomere FISH and FLAG immunostaining. Merge: telomeres (green), FLAG-POT1 (red), nuclei (blue). (B) Localization of γH2AX on metaphase spreads. Telomeres were detected by FISH, γH2AX by immunostaining. Merge: Merge: telomeres (green), γH2AX (red), chromosomes (blue). (C) Quantification of chromosomes with γH2AX foci on chromosome arms (Non-Tel), on one or more telomeres with detectable telomeric DNA (Tel) or at signal free ends (SFE). n = 3 independent experiments, mean ± S.E.M. (D) ChIP analysis showing changes in RPA and γH2AX localization at telomeres after CTC1 loss or POT1 overexpression. Chromatin from the indicated cell lines was precipitated with antibody to RPA or γH2AX. n = >3 experiments for each antibody, mean ± S.E.M.