Extended Data Figure 6.

Partial inhibition of mTor activity results in ES cell pausing. a, Representative image showing morphology of ES cells paused in 2i medium, with removal of the mTor inhibitor at passaging. Under these conditions, ES cells can be discontinuously paused in 2i medium. b, Western blot showing moderately reduced 4EBP1 and S6K1 phosphorylation, which are mediated by mTORC1, and abolished Akt phosphorylation, which is mediated by mTORC2, in paused ES cells. For gel source data, see Supplementary Figure 1. c, Flow cytometry analysis of nascent translation in the three states measured by HPG incorporation in E14 cells, in triplicates. d, Analysis of cell diameter in the three states, with each data point representing a population average of at least 300 cells per measurement. Values are represented as mean±SD. P-values are from two-tailed unpaired student's t-test. *: <0.05, **: <0.01, ***: <0.001, n.s.: not significant. e, Live pups born as a result of mating highly chimeric males generated using paused G4 ES cells with wild-type CD-1(ICR) female mice.