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- W2893198166 abstract "The ground state of pluripotency is defined as a minimal unrestricted epigenetic state as present in the Inner Cell Mass. Mouse embryonic stem cells (ESCs) grown in a defined serum-free medium with two kinase inhibitors (2i ESCs) have been postulated to reflect ground-state pluripotency, whereas ESCs grown in the presence of serum (serum ESCs) share more similarities with post-implantation epiblast cells. Pluripotency results from an intricate interplay between cytoplasmic, nuclear and chromatin-associated proteins. Here, we perform quantitative subcellular proteomics to gain insight in the molecular mechanisms sustaining the pluripotent states reflected by 2i and serum ESCs. We describe a full SILAC workflow and quality controls for proteomic comparison of 2i and serum ESCs, allowing subcellular proteomics of the cytoplasm, nucleoplasm and chromatin. The obtained quantitative information revealed increased levels of naïve pluripotency factors on the chromatin of 2i ESCs. Surprisingly, the cytoplasmic proteome suggests that 2i and serum ESCs utilize distinct metabolic programs, which include upregulation of free radical buffering by the glutathione pathway in 2i ESCs. Through induction of intracellular radicals, we show that the altered metabolic environment renders 2i ESCs less sensitive to oxidative stress. Altogether, this work provides novel insights into the proteomic landscape underlying ground state pluripotency." @default.
- W2893198166 created "2018-10-05" @default.
- W2893198166 creator A5007671569 @default.
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- W2893198166 date "2018-12-01" @default.
- W2893198166 modified "2023-09-24" @default.
- W2893198166 title "Quantitative subcellular proteomics using SILAC reveals enhanced metabolic buffering in the pluripotent ground state" @default.
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- W2893198166 doi "https://doi.org/10.1016/j.scr.2018.09.017" @default.
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