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. 2011 Mar 9;20(11):2171–2181. doi: 10.1093/hmg/ddr103

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© The Author 2011. Published by Oxford University Press

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.5), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 4. — USP1 depletion does not affect the repair of UVC-induced DNA lesions and does not improve FANCD2 foci formation. (A) HeLa cells transfected with Ctrl or USP1 siRNAs were treated with UVC (10 J/m²), and genomic DNA was prepared at the indicated times. Dot blots were performed with antibodies against 6-4 PP. DNA was then stained with an antibody recognizing single-stranded DNA. (B) Immunofluorescence analysis of the γH2AX (green) and FANCD2 (red) foci in Ctrl and USP1-siRNA-transfected cells treated with MMC (200 ng/ml for 24 h). Nuclei were stained with DAPI. Right images represent the merge of the three colours. (C) Quantification of cells with more than five FANCD2 nuclear foci in control- and USP1-siRNA-transfected cells left untreated and/or exposed to MMC (200 ng/ml) and fixed 24 h post-treatment. Data represent the mean of three independent experiments, and at least 100 cells were scored for each experiment.