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Figure 5. Depletion of RanBP1 decreases RCC1 dynamics. (a) FRAP analysis of RCC1 dynamics on metaphase chromatin in HCT116RanBP1-μAID/HA cells expressing RCC1-iRFP670::TIR1 and RanBP1-µAID-HA treated with (blue) or without (black) 1 mM Auxin for 3 h. (b) FLIP analysis cells, as in (A). (c) FRAP analysis of RCC1 dynamics on chromatin during anaphase B/telophase. Results are expressed as an average from a total of 12 measurements recorded from 3 independent experiments. (significance was assessed by F test). (d) RanBP1 was immunoprecipitated from mitotic lysates of control or Auxin-treated HCT116RanBP1-μAID/HA cells that express RanBP1-µAID-HA and RCC1-iRFP670 using HA antibody. Proteins were separated by SDS-PAGE and analyzed by immunoblotting with antibodies against RCC1, RanBP1, Ran, HA and RanBP3. Total lysate of parental HCT116 cells is shown in the first lane, for comparison purposes. Note both efficient interaction of RCC1 and RanBP1 (lane 4) and its specificity (lane 5). Image published in: Yau KC et al. (2020) This work was authored as part of the Contributor's official duties as an Employee of the United States Government and is therefore a work of the United States Government. Creative Commons Attribution-NonCommercial-NoDerivatives license Larger Image Printer Friendly View

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