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Figure 4


Fig. 4. Effects of APC overexpression on ß-catenin-induced ERK and ELK1 reporter activation. (A) DLD-1 cells were transfected with 0.5 µg of pcDNA3.0 or Flag-ß-catenin-pcDNA3.0 with or without 0.5 µg of pCMV-APC. They were harvested 72 hours later and western blot analyses were performed to detect p-ERK, Flag-ß-catenin, APC and ERK. Anti-Flag antibody was used to detect Flag-ß-catenin. (B) DLD-1 cells were transfected with 0.5 µg of pCMV or pCMV-APC with or without Flag-ß-catenin-pcDNA3.0. (Left panel) Co-transfection of 0.5 µg of ELK1 with 0.5 µg of pFR-Luc and 25 ng of pFA2-ELK1; ELK1 trans-reporter activity was measured. (Right panel) pTOPFLASH, pFOPFLASH were co-transfected, and luciferase activities derived from the reporter genes were measured 48 hours later. Wnt/ß-catenin reporter activity was calculated by dividing the luciferase activity of pTOPFLASH by that of pFOPFLASH (Korinek et al., 1997).





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