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. 2019 Nov 28;48(3):1285–1300. doi: 10.1093/nar/gkz1114

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© The Author(s) 2019. 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/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 2. — CSB is recruited to telomeric R-loops and contributes to cell survival and telomeric integrity. (A) Colony formation assays for U2OS and U2OS CSB KO cells with transiently expressing KR-TRF1 or RFP-TRF1. n = 3, error bars represent SD. (B) Telomere aberrations were found in U2OS and U2OS CSB KO cells with or without transiently expressing KR-TRF1. (C) Recruitment of CSB in KR-TRF1 or RFP-TRF1 expressing U2OS cells with or without light activation. Quantification of co-location frequency of myc-CSB and KR-TRF1/RFP-TRF1. (D) CSB foci co-localization with KR-TRF1 in U2OS cells treated with KU55933 (ATM inhibitor, 10 uM, 24 h), Nu7026 (DNA-PK inhibitor, 20 uM, 24 h), Olaparib (10 uM, 24 h), XAV-933(Tankyrase inhibitor,20 uM, 24 h), ATR inhibitor (10 uM, 24 h), and CDK2 inhibitor (5 uM, 24 h). (E) The staining of CSB at KR-TRF1 in U2OS cells after treating with overexpressed with HA-RNaseH WT, D210N mutant, DRB (20 μM, 24 h) and α-amanitin (100 μg/ml, 2 h), or TERRA knockdown by LNA. (F) CSB foci co-localization with KR-TRF1 in U2OS cells treated with TRF2 knockdown. (G) The staining of S9.6 at KR-TRF1 in U2OS cells after treating with CSB knockdown by siRNA. For (D)–(H), mean values with SD from 50 cells in three independent experiments are given. P-value is calculated by unpaired t-test, **P < 0.01, ***P < 0.001.