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. 2023 Jul 5;11:981425. doi: 10.3389/fcell.2023.981425

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Copyright © 2023 Didaskalou, Efstathiou, Galtsidis, Kesisova, Halavatyi, Elmali, Tsolou, Girod and Koffa.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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HURP interacting partners during mitosis (A) Co-immunoprecipitation assays using an antibody against HURP in HeLa Kyoto mitotic cell extracts. Rabbit IgG or anti-GFP rabbit polyclonal antibody were used as the respective controls. HURP and bound complexes were analyzed by Western blot using antibodies against HURP, TPX2 and two antibodies against Aurora A, as indicated on the left side (α-AurA antibody, or α-pAurA antibody). Input represents 6% of the total amount of protein used for the immunoprecipitation assay for HURP and p-Aurora A, 2% for TPX2, 5% for Aurora A. (B) Co-immunoprecipitation assay using an antibody against HURP in HeLa Kyoto mitotic cell extracts. Anti-GFP rabbit polyclonal antibody was used as a control. HURP and bound complexes were analyzed by Western blot using antibodies against HURP and Dynein. Input represents 2.5% of the total amount of protein used for the immunoprecipitation assay. (C) HeLa Kyoto mitotic extracts were immunoprecipitated using an antibody against HURP. The isolated complexes were analyzed by LC-MS/MS. Kif5B was identified as a possible partner of HURP. (D) Co-immunoprecipitation assays using either an antibody against the middle part (GKAP) or the N-terminal part of HURP (DHL5), in HeLa Kyoto mitotic cell extracts. Pre-immune IgG or Rabbit IgG were used as the respective controls. HURP and bound complexes were analyzed by Western blot using antibodies against HURP and Kif5B. Input represents 5% of the total amount of protein used for the immunoprecipitation assay.