Figure 4

Effects of NF-κB p65 siRNA on expression of MUC2. SEG-1 cells were transfected with Nuclear factor-κB (NF-κB) p65 siRNA, and its effect on NF-κB p65 mRNA was analyzed using (a) RT-PCR, and (b) Western blotting. (c) Cells were treated with 100 μM DCA for 18 hours after exposure to siRNA targeting NF-κB p65, (Upper panel) SEG-1 cells lysates were analyzed in Western blotting for MUC2 and β-actin, (Middle panel) The isolated RNA samples were analyzed by RT-PCR for MUC2, NF-κB p65 and β-actin, (Lower panel) SEG-1 cells were transfected with human MUC2 promoter luciferase construct, then treated for 18 hours with or without 100 μM DCA after with or without exposure to siRNA targeting NF-κB p65. Luciferase activity for MUC2 was measured and normalized to beta-galactosidase activity. (means ± SD of triplicate assays, * p < 0.05, for NF-κB p65 siRNA compared with assays without NF-κB p65 siRNA).