Figure 1. CSB does not affect the recruitment of OGG1 to oxidative DNA damage. (A) Representative stills of time-lapse ... Figure 1. CSB does not affect.

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Figure 1. CSB does not affect the recruitment of OGG1 to oxidative DNA damage. (A) Representative stills of time-lapse ... Figure 1. CSB does not affect the recruitment of OGG1 to oxidative DNA damage. (A) Representative stills of time-lapse imaging to determine accumulation kinetics of OGG1-GFP in CSB-proficient (upper panel) or CSB-deficient (lower panel) MRC-5 cells at micro-irradiated (405 nm) subnuclear regions, indicated by arrows, in the presence of 10 μM Ro 19-8022 photosensitizer. Scale bar: 7.5 μm. (B) Accumulation kinetics of OGG1-GFP at micro-irradiated areas (as shown in A). The mean relative fluorescence intensity is plotted against time in seconds. (C) FRAP analysis of OGG1-GFP expressed in CSB-proficient and deficient MRC-5 cells at locally damaged areas (on damage) or at a similar sized area without DNA damage (no damage). For (B) and (C), error bars indicate the SEM of 20 cells of 2 independent, pooled experiments. Unless provided in the caption above, the following copyright applies to the content of this slide: © The Author(s) 2018. 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 Non-Commercial 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 Nucleic Acids Res, Volume 46, Issue 15, 28 June 2018, Pages 7747–7756, https://doi.org/10.1093/nar/gky579 The content of this slide may be subject to copyright: please see the slide notes for details.

Figure 2. CSB facilitates recruitment of XRCC1 to BER-induced single strand breaks. (A) Representative stills of ... Figure 2. CSB facilitates recruitment of XRCC1 to BER-induced single strand breaks. (A) Representative stills of time-lapse imaging to determine accumulation kinetics of XRCC1-YFP in CSB-proficient or CSB-deficient MRC-5 cells on direct SSBs (405 nm laser micro-irradiated regions, indicated by arrows) in the absence (upper two panels) or presence of 15 μM PJ-34 to inhibit PARPs (lower two panels). (B) Accumulation kinetics of XRCC1-YFP at micro-irradiated areas (as shown in A). The mean relative fluorescence intensity is plotted against time in seconds. (C) Representative stills of time-lapse imaging to determine accumulation kinetics of XRCC1-YFP in CSB-proficient (upper panel) or CSB-deficient (lower panel) MRC-5 cells on BER induced SSBs cells at micro-irradiated (405 nm laser) subnuclear regions, indicated by arrows, in the presence of 10 μM Ro 19-8022 photosensitizer and 15 μM PJ-34. (D) Accumulation kinetics of XRCC1-YFP on BER induced SSBs (as shown in C). The mean relative fluorescence intensity is plotted against time in seconds. (E) Representative stills of time-lapse imaging to determine accumulation kinetics of the XRCC1<sup>L360D</sup>-YFP mutant in CSB-proficient (upper panel) or CSB-deficient (lower panel) MRC-5 cells at micro-irradiated (405 nm laser) subnuclear regions, indicated by arrows. (F) Accumulation kinetics of the XRCC1<sup>L360D</sup>-YFP mutant at micro-irradiated areas (as shown in E). The mean relative fluorescence intensity is plotted against time in seconds. (G) Representative stills of time-lapse imaging to determine accumulation kinetics of the XRCC1<sup>L360D</sup>-YFP mutant in CSB-proficient (upper panel) or CSB-deficient (lower panel) MRC-5 cells at micro-irradiated (405 nm laser) subnuclear regions, indicated by arrows, in the presence of 10 μM Ro 19-8022 photosensitizer. (H) Accumulation kinetics of the XRCC1<sup>L360D</sup>-YFP mutant at micro-irradiated areas (as shown in G). The mean relative fluorescence intensity is plotted against time in seconds. For (A, C, E and F), scale bar: 7.5 μm. For (B, D, F and H), error bars indicate the SEM of 20 cells of two independent, pooled experiments. Unless provided in the caption above, the following copyright applies to the content of this slide: © The Author(s) 2018. 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 Non-Commercial 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 Nucleic Acids Res, Volume 46, Issue 15, 28 June 2018, Pages 7747–7756, https://doi.org/10.1093/nar/gky579 The content of this slide may be subject to copyright: please see the slide notes for details.

Figure 3. XRCC1 binding to BER induced breaks is sensitive to inhibition of transcription. (A) Representative stills of ... Figure 3. XRCC1 binding to BER induced breaks is sensitive to inhibition of transcription. (A) Representative stills of time-lapse imaging to determine accumulation kinetics of XRCC1<sup>L360D</sup>-YFP in untreated (upper panel) or transcription inhibited (DRB, 100 μM) MRC-5 cells at micro-irradiated (405 nm laser) subnuclear regions, indicated by arrows, in the presence of 10 μM Ro 19-8022 photosensitizer. Scale bar: 7.5 μm. (B) Accumulation kinetics of XRCC1<sup>L360D</sup>-YFP at micro-irradiated areas (as shown in A). The mean relative fluorescence intensity is plotted against time in seconds. Error bars indicate the SEM of 20 cells of 2 independent, pooled experiments. (C) Accumulation kinetics of XRCC1<sup>L360D</sup>-YFP expressed in HeLa cells at micro-irradiated areas in the presence of 5 μM Ro 19-8022 photosensitizer in either untreated cells or in the presence of transcription inhibitor (DRB, 100 μM). (D) Accumulation kinetics of OGG1-dsRed expressed in HeLa cells at micro-irradiated areas in the presence of 5 μM Ro 19-8022 photosensitizer in either untreated cells or in the presence of transcription inhibition (DRB, 100 μM). All graphs are representative for two independent experiments (at least 11 cells) and show the mean relative fluorescence intensity ± SEM. Unless provided in the caption above, the following copyright applies to the content of this slide: © The Author(s) 2018. 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 Non-Commercial 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 Nucleic Acids Res, Volume 46, Issue 15, 28 June 2018, Pages 7747–7756, https://doi.org/10.1093/nar/gky579 The content of this slide may be subject to copyright: please see the slide notes for details.

Figure 4. Proposed model for the CSB function in the repair of oxidative DNA damage. The upper panel represents the ... Figure 4. Proposed model for the CSB function in the repair of oxidative DNA damage. The upper panel represents the classical BER initiation on 8-oxoG lesions (red star) by the concerted action of the OOG1 glycosylase (orange sphere) and APE1 (yellow ball), subsequent recruitment of the XRCC1/LigaseIII complex (blue and red balls, respectively) and DNA Polymerase β (Pol β, dark purple ball) through interaction with the glycosylase further completes repair. The lower panels represent a situation in which an elongating RNA polymerase II complex (RNAPII, purple sphere, direction indicated by curved arrow) runs into a BER-intermediate (e.g. BER-generated SSB) that likely caused inhibition of RNAPII elongation. When CSB (green ball) is present (lower left panel, blue shaded) it will assist in loading the XRCC1/Ligase III (LigIII) complex (blue and red balls, respectively) to stalled RNAPII on BER-intermediates to facilitate the assembly of downstream BER factors, such as DNA Polymerase β (Pol β, dark-purple ball) and stimulate BER progression. In the absence of CSB (lower right panel, orange shaded), BER progression will be limited. Unless provided in the caption above, the following copyright applies to the content of this slide: © The Author(s) 2018. 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 Non-Commercial 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 Nucleic Acids Res, Volume 46, Issue 15, 28 June 2018, Pages 7747–7756, https://doi.org/10.1093/nar/gky579 The content of this slide may be subject to copyright: please see the slide notes for details.