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. Author manuscript; available in PMC: 2013 Dec 14.
Published in final edited form as: Mol Cell. 2012 Oct 30;48(5):723–733. doi: 10.1016/j.molcel.2012.09.026

Figure 5. H2A.Z is required to retain the Ku70/80 complex at DSBs.

Figure 5

(a) Upper panel: U2OS cells expressing shCon or shH2AZ were irradiated (2Gy/60mins), fixed and RPA32 detected by immunofluorescent staining. Lower panel: U2OS cells expressing shCon or shH2AZ were prelabeled with BrdU then treated with camptothecin (CPT: 1μM) for 60 mins. Cells were fixed under non-denaturing conditions and BrdU in ssDNA detected with antiBrdU antibody. (b) Quantitation of (a). Cells with > 5 foci were counted as positive, with an average of 100 cells counted per experiment. p-values calculated using a t-test. (c) Laser striping was used to create DNA damage in U2OS cells expressing shCon or shH2AZ. Cells were pre-extracted (see methods) immediately after completion of laser striping (0 minutes) or allowed to recover for 15 or 30 minutes. γH2AX and Ku70/80 were detected by immunofluorescent staining. See figure S6 and S7.