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

. 2024 Mar 12;15:2210. doi: 10.1038/s41467-024-46578-0

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

© The Author(s) 2024

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

PMC Copyright notice

Fig. 1 — A Schematic of ZNF827 domains and functional mutants. B Electrophoretic mobility shift assay (EMSA) using γ-³²P radiolabelled single-stranded and double-stranded pentaprobes with increasing concentrations of purified ZNF827 protein (n = 1). Bound complex is indicated by the gel shift. C Representative image of ZNF827 (green) and native BrdU-stained ssDNA (red) colocalizations in U-2 OS cell nuclei following transient overexpression of ZNF827 (left panel). White arrows indicate colocalizations. Scale bar represents 5 μm. Manual quantitation of ZNF827 and ssDNA foci per nuclei and the fraction of colocalizing foci per nuclei (right panels). Data represent 50 nuclei from three biological replicates. D EMSA using γ-³²P radiolabelled single-stranded and double-stranded pentaprobes with purified ZNF827 wild-type and ZNF827 ZnF mutant proteins (n = 1). Bound complex is indicated by the gel shift. E Western blot analysis of RPA components, RPA32 and RPA70, and NuRD components, RBBP4 and HDAC1, following co-immunoprecipitation (co-IP) with a direct ZNF827 antibody in U-2 OS cells overexpressing ZNF827 (n = 1). F Western blot analysis of RPA32 following co-IP with a Myc antibody in U-2 OS cells overexpressing Myc-tagged ZNF827 and ZNF827 ZnF mutants (n = 1). Empty vector included as a negative control. G Representative images of Myc-tagged ZNF827 and ZNF827 mutants (green) and RPA32 (red) immunofluorescence labelling in U-2 OS ZNF827 knockout cells 48 h after transient transfection with Myc-tagged ZNF827 constructs (left panel). White arrows indicate colocalizations. Scale bar represents 5 μm. Manual quantitation of ZNF827 foci colocalizing with RPA foci (right panel). Data represent 50 nuclei from three biological replicates and data from individual nuclei were plotted as Tukey box plots; ****P < 0.0001, *P = 0.05. Multiple comparisons were corrected for using the Bonferroni test. H EMSA using γ-³²P radiolabelled single-stranded pentaprobe with purified ZNF827 and RPA protein complex (n = 1). Z denotes addition of ZNF827 15 min before RPA. R denotes addition of RPA 15 min before ZNF827. Bound complex is indicated by the gel shift, and the associated schematic indicates possible protein binding configurations. Source data are provided as a Source Data file.