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. 2018 Aug 13;19(8):2376. doi: 10.3390/ijms19082376

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© 2018 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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Expression of eGFP-TopBP1 W1145R induces accumulation of single-stranded DNA (ssDNA), but no DNA damage response. (a) ssDNA analysis of eGFP-TopBP1 WT and W1145R cells left non-induced or induced for 24 or 48 h. Means of three independent experiments with standard deviations are shown; (b) Representative examples of nuclei from panel A. DNA replication foci were labeled with a short pulse of EdU; (c) Co-localization of DNA replication foci (yellow) and ssDNA is shown in the overlay image (EdU + ssDNA) and in a magnified region marked by white frame (bottom frame); (d) Immunoblot analysis of whole-cell extracts from cells induced to express either eGFP-TopBP1 WT or W1145R for the indicated times. Etoposide was used as a positive control to induce the intra-S-phase damage response (normal human osteosarcoma (U2OS) cells).

Expression of eGFP-TopBP1 W1145R induces accumulation of single-stranded DNA (ssDNA), but no DNA damage response. (a) ssDNA analysis of eGFP-TopBP1 WT and W1145R cells left non-induced or induced for 24 or 48 h. Means of three independent experiments with standard deviations are shown; (b) Representative examples of nuclei from panel A. DNA replication foci were labeled with a short pulse of EdU; (c) Co-localization of DNA replication foci (yellow) and ssDNA is shown in the overlay image (EdU + ssDNA) and in a magnified region marked by white frame (bottom frame); (d) Immunoblot analysis of whole-cell extracts from cells induced to express either eGFP-TopBP1 WT or W1145R for the indicated times. Etoposide was used as a positive control to induce the intra-S-phase damage response (normal human osteosarcoma (U2OS) cells).