Figure 4.

The crosstalk between HIF-1α and NF-κB signaling pathway. (A) HCC-LM3 and SMMC-7721 cells were treated with 200 μM CoCl₂ for 24 h and 48 h. HIF-1α, the targeted protein CA9, VEGF, NF-κB signaling downstream gene c-Myc and ICAM-1 were assessed by Western blotting. Quantification plots are shown on the right. (B) HCC-LM3 and SMMC-7721 cells were treated with 200 μM CoCl₂ for 24 h; HIF-1α, CA9, VEGF, cMyc and ICAM-1 mRNA levels were detected by RT-PCR. Gene expression is normalized to GAPDH. The data were presented as mean ± SD, * p < 0.05, ** p < 0.01 compared with untreated cells. (C) HCC-LM3 and SMMC-7721 cells were induced with 10 ng/mL and 20 ng/mL TNFα for 24 h; HIF-1α, CA9 and VEGF protein expression were determined by Western blotting. Quantification plots are shown on the right. (D) HCC-LM3 and SMMC-7721 cells were induced with 20 ng/mL TNFα for 24 h; HIF-1α, CA9 and VEGF mRNA expression values were assessed by RT-PCR. Gene expression is normalized to GAPDH. The data were presented as mean ± SD, * p < 0.05, ** p < 0.01 compared with control group. Data are representative of three independent experiments.