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. Author manuscript; available in PMC: 2014 Sep 8.
Published in final edited form as: Nat Cell Biol. 2011 Aug 14;13(9):1070–1075. doi: 10.1038/ncb2314

Figure 4.

Figure 4

LIF and Wnt3a are sufficient to support ESC self-renewal. (a,b) Expansion over multiple passages of alkaline phosphatase-positive (AP+) R1 ESC colonies in medium containing serum and LIF (a) or in N2B27 containing LIF (b) (mean + s.e.m., n = 3). (c,d) Alkaline phosphatase-stained R1 ESCs cultured on MEFs (c) or maintained for six passages in N2B27 medium supplemented with LIF and Wnt3a (d). (e) Gene expression profile of R1 ESCs following four passages in the indicated conditions. (f) Expansion of alkaline phosphatase-positive R1 ESC colonies in N2B27 medium. Where indicated, Wnt3a was added at 200 ng ml−1. (g) Expansion of alkaline phosphatase-positive R1 ESC colonies in N2B27 medium containing LIF and PD0325901. (h) MEK activity (indicated by the presence of phospho-ERK (p-ERK; extracellular signal-regulated kinase) on western blot) in R1 ESCs cultured in N2B27 with LIF, was repressed by PD0325901 (PD) but not by Wnt3a, or by withdrawal of Wnt3a for 9 h (−9 h). Uncropped images of blots are shown in Supplementary Fig. S4g. Scale bar, 200 μm (c,d).