Fig. 2. xStAx-VHLL inhibits Wnt/β-catenin signaling.

a HEK293T cells transfected with Topflash luciferase were treated with Wnt3a CM and peptides (70 μM each) for 24 h and then harvested for luciferase measurement. SA SAHPA1, SA-V SAHPA1-VHLL, xS xStAx, xS-V xStAx-VHLL. b HEK293T cells transfected with Topflash luciferase were treated with Wnt3a CM and xStAx (xS) or xStAx-VHLL (xS-V) at indicated concentration for 24 h and then harvested for luciferase activity measurement. c HEK293T cells transfected with Topflash luciferase were treated with Wnt3a CM and 10 μM XAV939 (XAV), 20 μM PRI-724 (PRI), 100 μM PNU-74654 (PNU), 10 μM MSAB or 70 μM xStAx-VHLL (xS-V) for 24 h and then harvested for luciferase activity measurement. d HEK293T cells were treated with MSAB (10 μM) or xStAx-VHLL (70 μM) and Wnt3a CM for 24 h and then harvested for immunoblotting. e–g HCT116 (e), SW480 (f), and LoVo (g) cells were treated with MSAB (10 μM) or xStAx-VHLL (50 μM) for 24 h and then harvested for immunoblotting. h Equal amount (1 × 10⁴) of LoVo cells was treated with the vehicle DMSO, 10 μM MSAB or 50 μM xStAx-VHLL for 6 days, and the cell number was counted each day. i HEK293T cells were treated with MSAB (10 μM) xStAx-VHLL (70 μM) for 24 h and then replaced with fresh medium for 24 or 48 h to wash out the drugs before harvested for immunoblotting. The relative band intensity was quantified with ImageJ and normalized to GAPDH. Data from three independent experiments are displayed as the mean ± SD by two-way ANOVA (a, b, c, h) or one-way ANOVA (d–g). *P < 0.05, **P < 0.01.