Figure 1.

PAR₄-induced β-catenin stabilization. (A). β-catenin levels. HEK293 cells were co-transfected with either 0.2 μg or 0.4 μg Par4 and 0.3 μg flag-β-catenin plasmids and treated with 200 µM AYPGKF for 4 and 5 h to achieve PAR₄ activation. Western blot analysis was performed on cell lysates to detect β-catenin using anti-flag antibody (1:1000) and normalized to β-actin (1:1000) for protein loading. The figure is representative of the assay performed in triplicate. (B). Lef/Tcf tran-scriptional activity. HEK293 cells were transiently co-transfected with 0.2 μg Par4, 0.075 μg TOPflash, 0.125 μg LEF, and 0.2 μg β-gal plasmids. Luciferase activity was normalized to β-gal for transfection efficiency. The mean of duplicates is shown for each treatment. The results are representative of experiments performed in triplicate.