Figure 5

Overexpression of FHC induced by TNF-α inhibited apoptosis signaling in serum-deprived cells. (A and B) Serum-deprived Hep3B and SMMC-7721 cells were cultured with or without TNF-α (10 ng/ml) and/or BAY11-7082 (10 μM). The expression of FHC was determined by RT-PCR and Western-blotting analysis. Hep3B (C) and SMMC-7721 (D) cells were transfected with siFHC or Vector and then serum-deprived cells were treated with or without TNF-α (10 ng/ml) for 24 h. Western blotting was performed with anti-FHC or anti-caspase-8 or anti-cleaved caspase-3 or GAPDH. (E and F) Cell viability was detected by MTT assay. (G and H) Cells were harvested and stained with Annexin-V-FITC and PI, apoptosis was measured by flow cytometry. Data are presented as the mean ± SEM from three independent experiments. **P < 0.01; Student’s t-test.