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. Author manuscript; available in PMC: 2021 Mar 1.
Published in final edited form as: Gastroenterology. 2019 Dec 17;158(4):1029–1043.e10. doi: 10.1053/j.gastro.2019.11.302

Figure 6. Endogenous expression in cell lines confirms increased signaling and reduced affinity for APC.

Figure 6.

(A) Introduction of the K335I and S37F mutation in SNU449 cells leads to a modest increase in total β-catenin protein levels. In HEK293 cells this is restricted to S37F clones. Numbers in brackets refer to the number of CTNNB1 alleles expressing that variant. HEK293 clones are homozygous in all cases. (B) β-catenin luciferase reporter assay of all available clones. WRE/MRE ratios are shown. Data are presented as mean ± SD. **,P<0.01; ***, P<0.001; ****, P<0.0001. Bottom half shows the K335I clones in comparison with the parental line. (C) IP of endogenous APC reveals reduced binding of K335I. Two independent APC antibodies were used. Vertical dashed lines indicate removed irrelevant lanes.