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. 2021 Sep 1;49(17):9851–9869. doi: 10.1093/nar/gkab756

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© The Author(s) 2021. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial License (https://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com

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Figure 6. — Plk1 co-immunoprecipitates with MTBP, Treslin and TopBP1. (A) Egg extracts were incubated with control (IP Ctrl) or Plk1 (IP Plk1) antibody coupled Sepharose beads. Immunoprecipitates and input extract were subjected to gel electrophoresis and western blot analysis using the indicated antibodies. (B) Chromatin fractions, 60 min after the addition of 2000 nuclei/μl in mock- or Plk1-depleted extracts, analyzed by western blot with indicated antibodies. (C) Chromatin fractions, isolated 60 min after the addition of 2000 nuclei/μl in egg extracts were treated or not with λ phosphatase and subjected to western blotting with indicated antibodies. (D) Sperm nuclei were incubated in egg extracts in the absence or presence of the ATR/ATM inhibitor caffeine (5 mM) and the Chk1 inhibitor AZD7762 (0.5 μM), reactions were stopped at indicated times, chromatin was purified and subjected to western blotting with indicated antibodies. (E) Sperm nuclei were incubated in egg extracts in the absence or presence of the Cdc7 inhibitor PHA767491 (50 μM), reactions were stopped at 60 min, chromatin was purified and subjected to western blotting with indicated antibodies.