Wnt/β-catenin signaling inhibition reduces the expression of miR-181. (A) HT29-APC and HT29-GAL cells were incubated with 100 μM of ZnCL2 for different time points (0 hours, 24 hours and 48 hours). Then qRT-PCR analysis of all four mature miR-181s was performed in these cells. (B, C) β-catenin siRNA and control siRNA was transfected in HuH7 and HuH1 cells using lipofectamine 2000. Western blotting analysis of β-catenin and β-actin (left panel) and qRT-PCR analysis of CTNNB1 (right panel) were performed (B). qRT-PCR analysis of all four mature miR-181s (C). All these experiments were done in triplicates and the level of miRNAs and genes were shown as mean ± standard deviation. * refers to p < 0.05.