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. 2014 Mar 25;9(3):e92381. doi: 10.1371/journal.pone.0092381

Figure 6. Treatment with exogenous CS-E represses Wnt/beta-catenin signaling in ES cells and EB cultures.

Figure 6

(A) Immunofluorescence detection of beta-catenin (green) in ES cells in the absence of LIF (nuclei are stained blue with DAPI). Cells cultured in the absence of LIF and treated with the control L-CM did not show any nuclear beta-catenin, but low levels of membrane-bound beta-catenin; treatment with either C4S or CS-E did not alter levels or distribution of beta-catenin. When cultures were stimulated with W3a-CM, we observed a strong increase in nuclear accumulation of beta-catenin (red arrows). This nuclear accumulation was not altered by treatment with C4S, but was completely abolished by treatment with CS-E. Scale bar = 5 micrometer. (B,C) TOPFLASH reporter assays. In both ES cells cultured in the absence of LIF (B) and EBs on d10 of differentiation (C), treatment with exogenous CS-E could significantly inhibit TOPFLASH reporter activity induced by W3a-CM, while treatment with exogenous C4S had no significant effect. (*: p<0.05).