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. 2012 May 8;13(6):561–568. doi: 10.1038/embor.2012.58

Figure 1.

Figure 1

NBS1 Ser 432 is a CDK substrate. (A) Biochemical kinase assay with recombinant CDK1/CyclinB and purified MRN as substrate. (B) Alignment of region surrounding putative phospho-site in NBS1 orthologues. (C) Characterization of the NBS1–Ser 432 phospho-specific antibody. (D) NBS1 phospho-Ser 432 antibody recognizes NBS1 in whole-cell extracts and (E) after immunoprecipitation. (F) U2OS cells were treated or mock-treated with 50 μM roscovitine for 4 h. (G) Depletion of CDK1 or CDK2 in U2OS cells reduces NBS1 Ser 432 phosphorylation. (H) NBS1 Ser 432 phosphorylation is cell-cycle regulated. (I) NBS1 Ser 432 phosphorylation is reduced after DNA-damage induction. CDK, cyclin-dependent kinase; IP, immunoprecipitation; IR, ionizing radiation; MRN, MRE11–RAD50–NBS1; NBS, Nijmegen breakage syndrome; siRNA, small-interfering RNA; WT, wild-type.