Figure 4.

(A) The effect of CMO on constitutive NF-κB DNA-binding activity. HepG2 cells (5 × 10⁵/ml) and HCCLM3 cells (5 × 10⁵/ml) were treated with CMO for 4, 8, and 12 h. Nuclear extracts were prepared, 50 µg of the nuclear extract protein was taken for DNA-binding assay as described in Section “Materials and Methods.” (B,C) CMO inhibits constitutive activation of reporter gene expression. HepG2 (5 × 10⁵/ml) and HCCLM3 (5 × 10⁵/ml) cells were transfected with NF-κB luciferase and β-galactosidase reporter plasmid using lipofectamine, incubated for 24 h, and then treated with CMO for 4, 8, and 12 h. Cells were lysed in reporter lysis buffer and analyzed for luciferase activity and normalized with β-galactosidase activity. Results are expressed as % fold activity over the activity of vector control. *p < 0.05.