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. 2014 Nov 1;25(21):3424–3436. doi: 10.1091/mbc.E14-04-0885

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© 2014 Kunttas-Tatli et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

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FIGURE 4: — APC2 self-association is necessary to degrade β-cat and regulate Wnt target gene expression in SW480 cells. (A) Similar to S2 cells, Axin-GFP forms cytoplasmic puncta (1) and mCh-APC2 colocalizes with Axin-GFP in SW480 cells (2). Coexpression of APC2-ΔASAD with Axin-GFP does not disrupt colocalization (3) but is also associated with defects in puncta assembly and morphology. (B) Similar to S2 cells, coexpression of Axin with APC2-FL in SW480 cells leads to fewer, larger puncta. Two-tailed z test demonstrates significant differences between the two conditions. (C, D) Expression of full-length Drosophila APC2 was sufficient to suppress the elevated levels of β-cat (2) (compare to the empty vector control [1]) in SW480 cells. The APC2-ΔASAD mutant moderately suppressed the elevated β-cat levels (3). (E) In SW480 cells, expression of APC2-FL strongly suppressed activation of Wnt targets as assessed by TOP/Flash activity compared with the empty vector control. Expression of the APC2-ΔASAD mutant suppressed target gene activation compared with the empty vector control but exhibited significantly less activity than APC2-FL. Student's t test revealed significant differences between the conditions in D and E. (F) mCh-tagged APC2-FL (2) and APC2-ΔASAD (3) were expressed at equal levels in SW480 cells used in the TOP/Flash assays. Scale bar, 10 μm.