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The study of SOX2 deposition in asynchronous and mitotic E14TG2a cells

ABSTRACT: We executed ChIPseq for SOX2 a different mouse strain E14TG2a.

ORGANISM(S): Mus musculus

PROVIDER: GSE216663 | GEO | 2023/03/19

REPOSITORIES: GEO

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The study of SOX2 deposition in asynchronous and mitotic E14TG2a mouse ES cells.

Project description:We executed CUT&RUN-seq for SOX2 a different mouse strain E14TG2a

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The study of SOX2 deposition in asynchronous and mitotic E14TG2a cells

Project description:The study of SOX2 deposition in asynchronous and mitotic E14TG2a cells

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The study of SOX2 deposition in asynchronous and mitotic E14TG2a mouse ES cells.

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The study of SWI/SNF subunits and other transcription factors deposition in asynchronous and mitotic mESCs and cells released from mitosis

Project description:We executed CUT&RUN-seq for SWI/SNF components ARID1A, BRD9, SMARCA4, SMARCB1, SMARCE1, as well as ESRRB, SOX2, and EZH2 in asynchronous and mitotic cells and reported that, in asynchronous cells, ARID1A localized primarily at enhancer regions and EZH2 preferentially deposited at bivalent promoters and silent enhancer domains. The remaining factors were enriched at both TSS/promoters and to varying degrees at active enhancers. Unexpectedly, in mitosis, the chromatin regulatory factors almost all tethered at proximal gene regions with very little binding at enhancers. While the SWI/SNF subunits were bound principally at promoters, EZH2, the catalytic subunit of Polycomb Repressive Complex 2 was bound at both promoters and silent enhancers in mitotic cells. Moreover, we reported that upon the degradation of SMARCE1 in mitosis, the occupancy of SOX2, ESRRB, and EZH2 on mitotic chromatin was disrupted.

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Project description:E14Tg2a mouse embryonic stem cells (ESCs) were cultured on gelatin coated plates in KO-DMEM medium supplemented with 15% FCS and 1000 U/ml leukaemia inhibitory factor. Rapidly dividing cultures of ESCs were arrested in metaphase using demecolcine (0.1 μg/ml for 6 h), achieving samples where most (85-90%) cells were in M-phase as judged by propidium iodide labelling. Condensed chromosomes were released using polyamine buffer, stained with Hoechst 33258 and Chromomycin A3 and purified by flow cytometry using a Becton Dickinson Influx equipped with specialised air-cooled lasers.

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