Fig. 4.

Effects of EC supplementation on HFD-induced alterations in ileum tight junction protein expression. Mice were fed a control diet (empty bars), the control diet supplemented with 20 mg EC/kg body weight (dashed bars), a HFD (HF) (black bars), or the HFD supplemented with 20 mg (HFE20) EC/kg body weight (grey bars). A- TJ proteins occludin, claudin-1 and ZO-1, expression was measured by Western blot. Bands were quantified and values referred to HSC70 levels (loading control). Results for HF, HFE20 and CE were referred to control group values (C). Results are shown as mean ± SE of 8 animals/treatment. B- Representative images for immunohistochemistry and confocal microscopy for occludin and ZO-1 (red fluorescence), and claudin-1 (green fluorescence). Hoechst was used for nuclear counterstaining (blue). Bar: 50 µm C- Fluorescence intensity was measured as described in methods and results are shown as mean ± SE of 4 animals/treatment. A,C- #Significantly different from all other groups; (p < 0.05, one way ANOVA test).