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. 2014 Mar 6;10(3):e1004178. doi: 10.1371/journal.pgen.1004178

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© 2014 Leung et al

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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(A) Maximum H2AX-K13/15ub levels (−) or (+) RNF168 is dependent on the acidic patch. HEK293T cells were transfected with H2AX and derivatives either (−) or (+) RNF168 and analyzed as in Figure 1D. (B) Human H2AX and derivatives reconstituted in MCF10A H2AX^−/− cells. Western blot analysis of the indicated MCF10A H2AX^−/− stable cell lines. (C) H2AXub is dispensable for 53BP1 foci formation after DNA damage. Reconstituted MCF10A H2AX^−/− cells stably expressing H2AX and H2AX mutants were analyzed by immunofluorescence (IF) with the indicated antibodies. Cells were treated with 3 Gy IR and analyzed by IF 2 h post-IR.