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. 2008 Apr 21;181(2):227–240. doi: 10.1083/jcb.200709008

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Copyright © 2008, The Rockefeller University Press

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Figure 6. — Disruption of the MDC1–NBS1 interaction triggers a partial G2/M checkpoint defect. (A) NBS-iLB1 fibroblasts and NBS-iLB1 fibroblasts stably transduced with wild-type NBS1 and R28A mutant NBS1 were irradiated, fixed with methanol, and stained with antibodies against MDC1 and NBS1. (B) Whole cell extracts of NBS-iLB1 fibroblasts and NBS-iLB1 fibroblasts stably transduced with wild-type (WT) NBS1 and R28A mutant NBS1 were resolved by SDS-PAGE followed by immunoblotting. The blot was probed with antibodies against NBS1 and tubulin. (C) NBS-iLB1 fibroblasts and NBS-iLB1 fibroblasts stably transduced with wild-type NBS1 and R28A mutant NBS1 were left untreated or irradiated with 1 Gy and 3 Gy, respectively. Cells were harvested 1 h after irradiation, fixed with methanol, and stained with an antibody against phosphorylated H3 (P-H3) and propidium iodine. The percentage of phosphorylated H3–positive cells was determined by FACS analysis. Error bars represent SD. Bar, 10 μm.