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

. 2007 Dec 31;179(7):1413–1426. doi: 10.1083/jcb.200706179

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

Copyright © 2007, The Rockefeller University Press

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/).

PMC Copyright notice

Figure 3. — PR-Set7 silencing causes massive DNA breaks during S phase, which leads to activation of the DNA damage checkpoint. (A) Representative comets of shLuc and shPR7 U2OS cells and shPR7 U2OS_SR cells. Alkaline comet analysis was performed 3 d after shRNA expression. At least 200 randomly chosen comets were captured. (B) Distribution of cells with different comet tail moments and quantification of the number of cells with comet tail moments greater than five (inset) are illustrated for each cell line. The tail moment and distribution were scored from 200 cells/slide by using Comet Imager 1.2.10 software. (C) Immunoblot analysis of the total levels of p53 and P21 and the levels of phosphorylation of ATM, ATR, CHK1, H2A.X, and p53 in U2OS (lanes 1 and 2) and U2OS_SR (lane 3) cells 3 d after either shLuc (lane 1) or shPR7 (lanes 2 and 3) expression. (D) Immunofluorescence of BrdU (a, c, and e) and γ-H2A.X (b, d, and f) in shLuc and shPR7 U2OS cells and shPR7 U2OS_SR cells 3 h after release from a thymidine block. (bottom) Quantification of double BrdU and γ-H2A.X–positive cells in shRNA-expressing U2OS and U2OS_SR cell lines. Cells were harvested and analyzed by immunofluorescence at the times indicated. The percentages of double BrdU and γ-H2A.X–positive cells were obtained by counting 300 cells from three independent experiments. Error bars represent SD. (E) Images of asynchronous shPR7 U2OS nuclei acquired with Apotome microscopy 3 d after shRNA expression. After BrdU incorporation for 10 min, cells were stained with anti-BrdU antibody to detect active replication foci (a) and anti-53BP1 antibody to detect DNA damage foci (b). The merged picture (c) showed a partial but significant overlay. Bars, 5 μm.