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. 2011 Sep 20;40(2):701–711. doi: 10.1093/nar/gkr744

Figure 6.

Figure 6.

Cells from Ubr3-knockout mice are genetically instable. (A and C) MEFs from wild-type and Ubr3-knockout mice were fixed and immunostained with the indicated antibodies and stained with DAPI. (B) Graphic presentation of nuclei size distribution in wild-type and Ubr3-knockout fibroblasts (500 cell nuclei were evaluated). (D) Graphic representation of 53BP1 foci ratios calculated from three individual slides (53BP1 foci were counted from 100 cells/slide). (E) Whole-cell extracts from MEFs from wild-type (WT) and Ubr3-knockout (KO) mice were prepared and analysed by 10% SDS–PAGE and immunoblotting using the indicated antibodies. (F) HeLa cells were grown on 10-cm2 dishes for 24 h to 70–80% confluency and then transfected with human Flag-tagged APE1 (APE1Flag) expression plasmid. After 24 h cells were pelleted by centrifugation, whole cell extracts were prepared and analysed by 10% SDS–PAGE and western blotting with the antibodies indicated. (G) MEFs from wild-type and Ubr3-knockout mice were grown on 10-cm2 dishes in triplicate and treated with the indicated concentrations of MMS for 45 min and colonies allowed to grow for 7 days. The graph represents the mean surviving fraction normalized against the untreated controls from three independent experiments and the standard errors are shown.