Figure 5. Exogenously expression of p65 abrogated the effect of solamargine on MUC1 expression.

(A) PC3 and DU145 cells ells were exposed to increased concentration of solamargine (6 μM) for 24 h. Afterwards, the expression of p65 and p50 proteins was detected by Western blot. (B) PC3 and DU145 cells were treated with compound C (10 μM) for 2 h before exposure of the cells to solamargine for an additional 2 h. Afterwards, the expression of p65 protein were detected by Western blot. (C) PC3 and DU145 cells were transfected with the control or AMPKα siRNA (50 nM) for 24 h before exposing the cells to solamargine for an additional 24 h. Afterwards, AMPKα and p65 protein levels were determined by Western blot. (D) PC3 and DU145 cells were transfected with the control (pCMV4) or expression construct of p65 for 24 h before exposing the cells to solamargine for an additional 24 h. Afterwards, p65 and MUC1 protein expression were determined using Western blot. (E) PC3 and DU145 cells were transfected with the control (pCMV4) or expression construct of p65, and wild type human MUC1 promoter reporter construct ligated to luciferase reporter gene and internal control secreted alkaline phosphatase for 24 h before exposing the cells to solamargine (6 μM) for an additional 24 h. Afterwards, the Luciferase reporter activity was measured using Luciferase Assay System as described in the Materials and Methods Section. The figures are representative cropped gels/blots that have been run under the same experimental conditions. Values in bar graphs were given as the mean ± SD from three independent experiments performed in triplicate. *Indicates significant difference as compared to the untreated control group (P < 0.05). **Indicates significant difference from solamargine treated alone (P < 0.05).