Placzek, SiljaVanzan, LudovicaDeluz, CédricSuter, David M.2025-04-072025-04-072025-04-042025-04-2210.1016/j.celrep.2025.1154862-s2.0-105001054804https://infoscience.epfl.ch/handle/20.500.14299/248693Cell identity maintenance faces many challenges during mitosis, as most DNA-binding proteins are evicted from DNA and transcription is virtually abolished. How cells maintain their identity through division and faithfully re-initiate gene expression during mitotic exit is unclear. Here, we develop a novel reporter system enabling cell cycle synchronization-free separation of pluripotent stem cells in temporal bins of <30 min during mitotic exit. This allows us to quantify genome-wide reactivation of transcription, sequential changes in chromatin accessibility and transcription factor footprints, and re-binding of the pluripotency transcription factors OCT4, SOX2, and NANOG (OSN). We find that transcriptional activity progressively ramps up after mitosis and that OSN rapidly reoccupy the genome during the anaphase-telophase transition. We also demonstrate transcription factor-specific, dynamic relocation patterns and a hierarchical reorganization of the OSN binding landscape governed by OCT4 and SOX2. Our study sheds light on the dynamic orchestration of transcriptional reactivation after mitosis.trueATAC-seqChIP-seqchromatinCP: Cell biologyCP: Stem cell researchmitosisNANOGOCT4pluripotent stem cellsRNA-seqSOX2transcription factorstext::journal::journal article::research article