Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: UBR5-mediated ubiquitination of ATMIN is required for ionizing radiation-induced ATM signaling and function

doi: 10.1073/pnas.1400230111

Figure Lengend Snippet: ATMIN ubiquitination is required for ATM signaling and checkpoint function post IR. (A) G2/M nocodazole trap and quantification of mitotic index in 293T cells transfected with vector (CTR), Flag-ATMIN wild type, or Flag-ATMIN K238R, measured by FACS of phospho-histone 3 (pH3)-positive cells. (B) 53BP1 and pATM immunofluorescence staining of ATMINf/f MEFs transfected with empty vector (ATMINf/f+CTR) or ATMINΔ/Δ MEFs reconstituted with either wild-type (+wtATMIN) or K238R mutant Flag-ATMIN (+ATMIN-K238R). (C) Quantification of 53BP1-positive cells (with at least six distinct foci) from the experiment in B. (D) Western blots of whole-cell lysates from MEFs treated as in B. (E) Radiosensitivity assay of reconstituted MEFs as in B, showing percentage of surviving colonies 7 d after IR. Error bars represent SEM (***P < 0.005, **P < 0.01, *P < 0.05).

Article Snippet: Fixed cells were stained with phospho-Histone 3 antibody and analyzed using a BD Biosciences FACScan.

Techniques: Transfection, Plasmid Preparation, Immunofluorescence, Staining, Mutagenesis, Western Blot