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Activation of Wnt signaling enhances self-renewal of mouse embryonic and neural stem/progenitor cells. In contrast, undifferentiated ES cells show a very low level of endogenous Wnt signaling, and ectopic activation of Wnt signaling has been shown to block neuronal differentiation. Therefore, it remains unclear whether or not endogenous Wnt/ β -catenin signaling is necessary for self-renewal or neuronal differentiation of ES cells. To investigate this, we examined the expression profiles of Wnt signaling components. Expression levels of Wnts known to induce  β -catenin were very low in undifferentiated ES cells. Stable ES cell lines which can monitor endogenous activity of Wnt/ β -catenin signaling suggest that Wnt signaling was very low in undifferentiated ES cells, whereas it increased during embryonic body formation or neuronal differentiation. Interestingly, application of small molecules which can positively (BIO, GSK3 β  inhibitor) or negatively (IWR-1-endo, Axin stabilizer) control Wnt/ β -catenin signaling suggests that activation of that signaling at different time periods had differential effects on neuronal differentiation of 46C ES cells. Further, ChIP analysis suggested that  β -catenin/TCF1 complex directly regulated the expression of  Sox1  during neuronal differentiation. Overall, our data suggest that Wnt/ β -catenin signaling plays differential roles at different time points of neuronal differentiation.

Dual Function of Wnt Signaling during Neuronal Differentiation of Mouse Embryonic Stem Cells