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. Author manuscript; available in PMC: 2017 Feb 17.
Published in final edited form as: Mol Cell. 2015 Apr 30;58(5):780–793. doi: 10.1016/j.molcel.2015.04.001

Figure 4. β-Catenin Recruits Cohesin and Induces long-Range Enhancer-Promoter Looping in Wnt3a-Signaling hESCs.

Figure 4

(A) Heatmaps showing ChIP-seq read density of NIPBL and β-catenin at LEF-1 peaks in response to Wnt3a (200 ng/ml; 4 hr) in H1 hESCs.

(B) Coimmunoprecipitation of endogenous β-catenin and NIPBL proteins in untreated and Wnt3a-signaling hESCs.

(C) Genome browser image of the NIPBL and β-catenin binding profiles at the MIXL1 and EOMES loci in untreated and Wnt3a-treated hESCs. Below, 3C experiment was done in untreated or Wnt3a-treated hESCs. The diagram below each graph indicates the position of the primers (arrows indicate orientation) and the probe (green star). Blue and yellow boxes highlight upstream enhancers and the promoter, respectively. Mean (SD; n = 2).

(D) Graphs on the right show relative mRNA levels of several Wnt3a induced genes in NIPBL, SMC3 and MED26 siRNA transfected cells. The immunoblot on the left shows the knockdown efficiency for each siRNA. Mean (SD; n = 2–3).

See also Figure S4.