Effect of NC on the activation of SHH pathway in hepatic cancer xenograft mice and HepG2 cells. (A) Tumor tissues were processed for IHC for SHH and Gli-1. The photographs are representative images taken at a magnification of 400 ×. Quantification of IHC assay was represented as percentage of positively-stained cells. Data shown are averages with S.D. (error bars) from 6 individual mouse in each group. *P < 0.05; **P < 0.01, versus controls. (B) The protein expression levels of Shh and Gli-1 in HepG2 cells were determined by Western Blotting. β-actin was used as the internal control. The mRNA levels of SHH and Gli-1 in tumor tissues (C) and HepG2 cells (D) were determined by RT-PCR. GAPDH was used as the internal control. Images of Western Blotting and RT-PCR are representatives of 3 individual mouse in each group.