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. Author manuscript; available in PMC: 2013 May 11.
Published in final edited form as: Mol Cell. 2012 Mar 29;46(3):351–361. doi: 10.1016/j.molcel.2012.02.018

Figure 5. NBS1 Ubiquitination Is Critical for ATM Activation and Recruitment.

Figure 5

(A) 293T cells were transfected with indicated plasmids and harvested for in vivo ubiquitination assay.

(B) 293 cells transfected with Flag-NBS1 WT, K735R, and K751R respectively were treated with IR (3 Gy) and harvested for western blot analysis 5 min after IR treatment.

(C) 293 cells were transfected with Flag-NBS1 WT, K735R, and K751R, respectively, treated with or without IR (10 Gy), and harvested for the isolation of chromatin fraction 5 min after IR, followed by western blot analysis.

(D) HeLa cells transfected with WT-NBS1 and NBS1-K735R were treated with IR (3 Gy) and harvested for IF analysis 5 min after IR treatment. Percentages of cells with positive ATM-s1981p foci were quantified from cells with positive Flag-NBS1 transfection. Results are presented as mean values ± standard deviation (SD) from three different experiments.

(E) 293 cells were transfected with Flag-NBS1 WT and K735R, respectively, treated with IR (3 Gy), and harvested for coimmunoprecipitation assay at various time points, followed by western blot analysis.

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