The effects of aplykurodin A in SNU475 and Hep3B cells. (A) After incubation of TOPFlash and pCMV-Renilla luciferase (RL) plasmids co-transfected SNU475 and Hep3B cell with either DMSO or aplykurodin A (20 and 40 μM), FL and RL activities were determined. TOPFlash activity is normalized to RL activity. The results represent the mean ± S.D. of three independent experiments. * p < 0.01 compared with the DMSO control group. (B,C) After incubation of SNU475 and Hep3B cells with either DMSO or aplykurodin A (20 and 40 μM), cytosolic proteins were analyzed by Western blotting with anti-β-catenin and anti-β-actin antibodies (B) and cell extracts were immunoblotted with anti-cyclin D1, anti-c-myc, anti-Axin2 antibodies, and anti-β-actin (loading control) antibodies (C). The results are representative of three independent experiments. (D) The effect of aplykurdin A on cell viabilities of SNU475 and Hep3B cells. Details are described in Materials and Methods. The results represent the mean ± S.D. of three independent experiments. * p < 0.01 compared with the DMSO control group.