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. 2017 Feb 17;292(16):6461–6467. doi: 10.1074/jbc.M117.777474

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© 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

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Figure 1. — TIRR is 53BP1-associated protein. A, schematic presentation of 53BP1 and its IRIF region that is used in this study. B, 53BP1 IRIF region form foci following DNA damage. 293T cells stably expressing SFB-53BP1 IRIF region grown on coverslips were treated with 10 Gy of IR; cells were fixed, and immunostaining was carried out using indicated antibodies. C and D, lists of 53BP1 IRIF region or TIRR-associated proteins identified in soluble fraction by mass spectrometry analysis. E, endogenous 53BP1 interacts with TIRR. HeLa cell lysates were prepared and immunoprecipitated (IP) with TIRR antibody. Immunoprecipitates were blotted using antibodies as indicated. F, 53BP1 IRIF region specifically binds to TIRR. Beads coated with bacterially expressed GST-fused 53BP1 IRIF region were incubated with cell lysates containing exogenously expressed SFB-tagged TIRR. Immunoblotting experiments were carried out using the indicated antibodies. WB, Western blotting.