sFRP2 enhances: (A)Wnt3a-mediated β-catenin accumulation, (B) β-catenin nuclear translocation, (C) TCF/LEF transcriptional activity, and (D) LRP6 phosphorylation in HEK293A. Cells were treated with Wnt3a ± indicated sFRP2 for 2 h. The levels of total cellular and nuclear β-catenin as well as phosphorylated LRP6 were detected on Western blots. Antibodies against β-actin and TATA-binding protein (TBS) were used as loading controls. Shown are representative results of three independent experiments. For luciferase assays, cells were co-transfected with Renilla transfection-control plasmids plus either TopFlash (□) or FopFlash (■) plasmids. After transfection, cells were treated with Wnt3a ± sFRP2 for 24 h before extraction for luciferase assays. Shown are means ± SEM from at least 3 experiments each performed in triplicate wells. ***P<0.0001 compared with control by using of ANOVA and the Newman-Keuls test.