Figure 8.

Proteasome Inhibition Restores EZH2 Protein and Reduces Fibrosis in Mdr−/− Mice.

A. Western blotting on nuclear fractions of liver lysates from young (4 and 7 weeks) and old (14 and 40 weeks) Mdr2−/− mice show decreasing levels of EZH2 protein (left) with HSC70 as a loading control. Densitometry on the right shows a 3-fold decrease in EZH2 levels. *p<0.01. All error bars are SEM, n=4.

B. Mdr2−/− mice were treated with 1mg/kg Bortezomib from 7weeks to 11 weeks of age (males and females). Immunoblotting for EZH2 on nuclear fraction of liver lysates from both vehicle and Bortezomib treated mice show an increase in EZH2 protein levels with Bortezomib treatment (left), F: female, M: Male. Densitometry on the right shows 3-fold increase EZH2 protein when normalized to HSC70 as loading control. **p<0.001. All error bars are SEM, n=3.

C. RT-PCR analysis on the liver tissue from vehicle and Bortezomib treated Mdr2−/− mice show no change in EZH2 gene expression. All error bars are SEM, n=3.

D. Liver histology from Mdr2−/− mice treated with vehicle or Bortezomib show reduced fibrosis by Sirius red and Trichrome staining (left). Right shows the quantification for the % positive area for Sirius red and trichrome staining. ***p<0.0001. All error bars are SEM, n=4–5 animals per group.