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. 2012 Aug 21;23(1):122–130. doi: 10.1038/cr.2012.119

Figure 4.

Figure 4

DKK1 is a KDM6A/KDM6B target contributing to late endoderm differentiation. (A) RT-qPCR analysis of the expression of H3K27me3-enriched WNT antagonist genes during definitive endoderm differentiation. SFRP4 is not detectable. Expression levels are related to day 0. Data presented is the average of three independent experiments with error bars. (B) RT-qPCR analysis demonstrates downregulation of DKK1 expression upon KDM6A or KDM6B knockdown. (C) ChIP analysis demonstrates increased H3K27me3 level, but not H3K4me3, at the DKK1 gene enhancer upon KDM6A or KDM6B knockdown. The analysis was performed at the beginning of the second stage of differentiation. (D) Activin A-induced endoderm differentiation is compromised by DKK1 knockdown, which can be rescued by addition of WNT antagonist Xav939 at late stage of differentiation. The differentiation process includes 2-day treatment of activin A and Wnt3a followed by another 2-day treatment with or without Xav939. The differentiation efficiency is measured by quantification of CD184-positive cell population after 4-day differentiation.