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. 2008 Oct 20;183(2):241–252. doi: 10.1083/jcb.200803173

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© 2008 Franchitto et al.

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.jcb.org/misc/terms.shtml). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/).

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Figure 2. — Accumulation of DSBs after replication arrest observed in the absence of WRN depends on the presence of the MUS81 endonuclease. (A) MUS81 down-regulation by transfection of MUS81 siRNAs in wild-type and WS cells. (B) MUS81 and/or WRN down-regulation by transfection of MUS81 and/or WRN siRNAs in HeLa cells was verified by immunoblotting 48 h after transfection. (C) Formation of DSBs in wild-type and WS fibroblasts transfected with MUS81 siRNAs was evaluated by PFGE after 6 h of treatment with 2 mM HU or 1 μM etoposide (Etop.). Data are presented as mean ± SE from three independent experiments. (D) Formation of DSBs in HeLa cells transfected with control (GFP), MUS81, and/or WRN siRNAs and treated with 2 mM HU for the indicated times was evaluated by PFGE. Data are presented as mean ± SE from three independent experiments. (E) Analysis of DSB accumulation by anti–γ-H2AX immunofluorescence in HeLa cells transfected with control (GFP), MUS81, and/or WRN siRNAs and treated with 2 mM HU for 6 h. Representative images are shown. The percentage of γ-H2AX–positive nuclei from three independent experiments is shown. Data are presented as mean ± SE. Bars, 10 μM.