Figure 3. Chronic liver injury in CDE-diet fed mice is associated with loss of TJ adhesion-molecules.

(A) RT-PCR analysis shows significant decrease in mRNA expression of claudins-3, 5, 7 and increase in occludin in CDE-diet for 4d as compared to mice on normal-diet. Rescue was seen in claudins-3, 5, 7 level whereas further increase in occludin at 14d on CDE-diet. (B) Gel showing PCR product for claudins-3, 5, 7 and occludin. (C) In WT liver, claudin-3 and occludin localizes to the membrane in a punctate manner. Localization of claudin-3 to the membrane along with its colocalization with occludin is reduced in liver of mice fed on CDE-diet for 4d. Occludin levels are more at this stage. At 14d, claudin-3 level reappears to the membrane and colocalizes with occludin which continues to show an increase. (D) Membrane localization of claudin-5 and phalloidin staining marking the hepatocyte cytoskeleton in WT-liver. Membrane localization of claudin-5 is lost in CDE-diet fed mice (4d) and phalloidin staining indicates increases steatosis. At day 14 on CDE, claudin-5 level reappears whereas phalloidin staining indicates improved hepatic architecture. (E) In WT-liver, claudin-7 localizes to the membrane in a punctate manner. Localization of claudin-7 to the membrane in liver of mice fed on CDE-diet for 4d shows reduced expression. At 14d on CDE-diet, claudin-7 level reappears to the membrane. (F) Representative WB of junctional proteins shows reduction in claudins-3, 5, 7 at 4d post CDE-diet administration which is rescued at 14d. Occludin levels are increased at both 4d and 14d. GAPDH shows protein loading. * denotes p=0.05