Overexpression of Sox2 can negatively regulate HAMP expression in Huh7 cells. (A) Huh7 cells were transfected with an increasing amount of plasmids expressing Myc-Sox2 (pcDNA3.1-Myc vector was used to balance the DNA to the same quantity in each group), and cell were harvested for RT-qPCR assay to determine HAMP expression and also cell lysates were analyzed with immunoblotting with the anti-Myc or anti-actin antibodies at 48 h post-transfection. (B-C) Huh7 cells were transfected with the plasmids expressing Myc-Sox2. Forty-eight hours later, cells were stimulated with IL-6 (B) or BMP-2 (C) for additional 6 hours. And cells were harvested for RNA extraction to determine HAMP expression and some cell lysates were immunoblotted with the anti-Myc antibody to analyze the Myc-Sox2 expression. The experiments were repeated for three times with similar results and data were shown as means ± SE. Statistical analyses were conducted using one-way ANOVA with Tukey’s multiple comparison test. Significant differences are indicated by *p < 0.05.