Fig. 2.

Tributyrin supplementation sustains tight junction protein expression and co-localization in the ileum during chronic ethanol (EtOH) feeding. Mice were allowed free access to EtOH (25 days, 32%) or pair-fed control diets as described in Materials and Methods. Tributyrin (5 mM) was added to liquid diet for the first 11 days (1 to 4% EtOH) and then increased to 10 mM for days 12 to 25 (5 and 6% EtOH; 5 to 10 mM). Occludin (red) and ZO-1 (green) were visualized by immunohistochemistry in sections of ileum frozen in OCT. Nuclei were counterstained with DAPI (blue). A selected area was cropped and enlarged. All images were acquired using a 40× objective. Images are representative of at least replicate images captured per mouse in 4 to 6 mice (pair-fed) or 6 mice (EtOH-fed).