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. 2015 Sep 17;10(9):e0138710. doi: 10.1371/journal.pone.0138710

Fig 2. Effect of apigenin intake on NF-κB activation in the dorsolateral prostate of TRAMP mice.

Fig 2

(A) Protein expression of NF-κB/p65 and NF-κB/p50 as determined by Western blot analysis. A significant decrease in NF-κB/p65 and NF-κB/p50 is observed after apigenin intake. Actin as loading control. (B) IHC of NF-κB/p65 in the dorsolateral prostate from control and apigenin-treated TRAMP mice. A marked decrease in the expression of NF-κB/p65 is observed after apigenin intake. (C) NF-κB/p65 labeling index was calculated as number of NF-κB/p65 positive cells x 100 / total number of cells counted under x40 magnification in four randomly selected areas in each sample. Values represent Mean + SE, **P<0.05, compared to control. (D) DNA binding assay by EMSA. Nuclear extracts were prepared, and electrophoretic mobility shift assay was performed to assess NF-κB DNA binding activity. A marked decrease in the DNA binding of NF-κB/p65 is observed after apigenin intake. Controls were as follows: #1, biotin-Epstein-Barr virus Nuclear Antigen (EBNA) control DNA; #2, biotin-EBNA control DNA + EBNA extract; #3, biotin-EBNA control DNA + EBNA extract + 20-fold molar excess of unlabeled EBNA DNA. Details are described in ‘materials and methods’ section.