Figure 1.

Hypoxia up-regulates ZFHX3 expression in HCC cells. A and B, HCC cell lines HepG2 and Huh-7 were cultured under hypoxia for the indicated times, and the expression of ZFHX3 and two regulators of hypoxia-induced angiogenesis, HIF1A and VEGFA, was detected by Western blotting and real-time PCR for protein (A) and mRNA (B), respectively. ZFHX3 band intensities were quantified and normalized to β-actin, and the results are shown below the ZFHX3 bands in A. C–F, HepG2 and Huh-7 cells were treated with the hypoxia-mimetic agent DFO for the indicated times at 50 μm (C and D) or the indicated concentrations for 24 h (E and F), and expression of the same set of molecules was analyzed as in A and B. Data are shown as mean ± S.D. Band intensity ratios below each lane of Western blottings in A, C, and E were the average from three independent experiments, and their scatter plots and statistical details are shown in Fig. S1 (S1K–S1M). *, p < 0.05. The statistical analysis of real-time PCR was based on three independent experiments (i.e. n = 3), and the value for each group in an experiment was the average of triplicates. *, p < 0.05; **, p < 0.01; ***, p < 0.001; ns, not significant.