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. Author manuscript; available in PMC: 2017 May 23.
Published in final edited form as: Nature. 2016 Nov 23;540(7631):119–123. doi: 10.1038/nature20578

Figure 3.

Figure 3

mTor inhibition induces pausing of ES cells. a, Representative bright field images of ES cells cultured in serum, 2i and paused conditions (media compositions listed). b, Expansion curves of serum, 2i and paused ES cells. Two biological replicates, each with three technical replicates, were performed. c, Flow cytometry analysis of apoptotic cell populations in serum, 2i and paused ES cells. Annexin V and Sytox Blue were used for early and late apoptosis. 8 technical replicates were performed. d, Expansion curves of ES cells showing the reversible nature of the paused state. Two biological replicates, each with three technical replicates, were performed. e, Cell cycle analysis of serum, 2i and paused ES cells, based on four technical replicates of each state. f, Flow cytometry showing dilution of incorporated EdU in serum and paused ES cells. 3 technical replicates are shown. g, Chimeric mice generated from G4 ES cells previously paused for 7 days and released from pausing prior to morula aggregations with albino host embryos.