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 Apr 14;11:1819. doi: 10.1038/s41467-020-15689-9

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

© The Author(s) 2020

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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, visit http://creativecommons.org/licenses/by/4.0/.

PMC Copyright notice

Fig. 7 — a Top: Scheme of the synchronization procedure. Bottom: Quantification of misaligned chromosomes in BRCA2-T207A stable clones transiently overexpressing the 3xFLAG-BUBR1-3D-RFP mutant (S670D, S676D and T680D). n indicates the total number of cells counted from two independent experiments. Unpaired two-tailed t-test was used to calculate statistical significance of differences, ns (non-significant)). b Representative images of the type of chromosome alignment observed in cells overexpressing 3xFLAG-BUBR1-3D-RFP quantified in (a); scale bar represents 10 µm. c Co-immunoprecipitation of endogenous PP2A with transiently overexpressed 3xFLAG-BUBR1-3D-RFP mutant from mitotic cell extracts of BRCA2 WT cells or cells bearing the S206C variant using rabbit anti-RFP antibody. Rabbit IgG was used as control for the BUBR1 immunoprecipitation. The immunocomplexes were resolved on 4–15% SDS-PAGE followed by WB using mouse anti-BUBR1 and PP2AC antibodies. The amount of PP2A co-immunoprecipitated with BUBR1-3D-RFP relative to the input levels of PP2A and the amount of immunoprecipitated BUBR1-3D-RFP is presented below the blot as mean ± SD from two independent experiments. The data are presented relative to the non-treated BRCA2 WT. Asterisk denotes a non-specific band (d). Top: Scheme of the synchronization procedure. Bottom: Quantification of cells with aberrant chromosomes segregation in BRCA2^−/− cells and in the clones stably expressing BRCA2 WT, S206C and T207A, as indicated. Two-way ANOVA test with Tukey’s multiple comparisons test was used to calculate statistical significance of differences (the p-values show the significant differences compared to WT; ns (non-significant)). n in (d) indicates the total number of cells counted for each clone from two (BRCA2^−/−, S206C and T207A) and five (BRCA2 WT) independent experiments. e Representative images of the type of aberrant chromosome segregation observed in the cells quantified in (d), CREST antibody is used as marker of centromere; nuclei are revealed with DAPI counterstaining. Scale bar represents 10 µm. Source data are available as a Source Data file.