Figure 3

Berberine treatment regulates signaling pathways in intestinal tumors of Apc ^min/+ mice. (A-B) Immunohistochemical analyses of tumors using antibodies against β-catenin and cyclinD1 from the distal small intestine of untreated and 0.1% berberine-treated mice were shown (400×). Scale bars, 50 μm. Data were quantified as mean percentage of positive cells for β-catenin in cytoplasm and/or nuclear and cyclinD1 at five randomly selected fields. (C-E) p-EGFR, p-ERK and p-Akt were also analyzed by immunohistochemical staining. Data were quantified as mean percentage of positive cells. (F) Nuclear protein extracts were prepared from tumors, and analyzed by Western blot analysis using anti-β-catenin antibody. Anti-histone H3 antibody was used as a nuclear protein loading control. Protein lysates were prepared and analyzed by Western blot analysis using anti-total EGFR and p-EGFR antibodies, and anti-β-actin antibody was used as a protein loading control. The protein band ratio was calculated by comparing the relative density of the protein band for β-catenin, total EGFR, and p-EGFR to that of internal control band from the same mouse. The average ratio in control was set as 100%, the fold changes of the ratio in treated mice were shown. Columns, means from at least six mice in each group; bars, standard deviation. *, P < 0.01, 0.1% berberine-treated vs untreated Apc ^min/+ mice.