Fig. 5

BZM aggravated MTX-induced inhibition of Wnt/β-Catenin signaling. LNCaP and 22RV1 cells were treated with vehicle, BZM (200 nM), MTX (1 μM) alone and combination (BZM, 200 nM; MTX, 1 μM) for 24 h. A, D. Whole cellular lysates were subjected to western blot with indicated antibodies. Actin served as endogenous protein loading control. B, C. Cytoplasmic (B) and nuclear (C) protein were isolated after treatment for 24 h. Expression of β-Catenin was determined by western blot. E. Real-time PCR assay of indicated genes expression. LNCaP(left) and 22RV1(right) cells were treated with vehicle, BZM, MTX alone or combination for 24 h. Data are presented as from three independent experiments and mean ± s.e.m. The asterisk indicates a significant difference compared to the vehicle control (one-way ANOVA, ***p < 0.001)