Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2020 Aug 1;2(3):zcaa013. doi: 10.1093/narcan/zcaa013

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© The Author(s) 2020. Published by Oxford University Press on behalf of NAR Cancer.

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

PMC Copyright notice

Figure 5. — BRCA2 suppresses XRCC1–POLQ–MRE11 complex formation and MMEJ. (A) XRCC1 foci formation after treatment of scr-U2OS or BRCA2-U2OS cells with 3 mM HU and/or 3 μM ATRi for 8 h where indicated. Cells were marked positive if they contained >10 foci. At least 50 cells were analyzed for each experiment. (B) Representative images of the PLA data related to (E). (C) Representative images of the PLA data related to (F). (D) Upper panel: western blot of POLQ-FLAG IP from scr-U2OS and BRCA2-U2OS cells with and without ATRi + HU treatment. Lower panel: western blot of endogenous XRCC1 IP from scr-U2OS and BRCA2-U2OS cells expressing POLQ-FLAG with and without ATRi + HU treatment. (E) XRCC1 localization to sites of replication stress in scr-U2OS or BRCA2-U2OS cells, as measured by BrdU-XRCC1 PLA. Asynchronous cells were pulsed with 10 μM BrdU for 15 min before treatment with 3 mM HU and/or 3 μM ATRi for the indicated times. At least 50 cells were analyzed for each experiment. (F) PLA between XRCC1 and the indicated factors in scr-U2OS and BRCA2-U2OS cells treated for 8 h with 3 mM HU and 3 μM ATRi. At least 50 cells were analyzed for each experiment. (G) MMEJ repair activity of XRCC1-FLAG IP. scr-U2OS and BRCA2-U2OS cells were transfected with XRCC1-FLAG and treated with 3 mM HU and 3 μM ATRi where indicated. Immunoprecipitated XRCC1-FLAG complexes were incubated with linearized plasmid substrate and competent bacterial cells were transformed with repaired product according to (30). Western blot of IP bead eluate after incubation is shown.