The Mbd1 protein was localized mainly in the hepatocyte nuclei and demonstrated stronger staining in hepatocytes from B6 Mdr2-KO mice versus Mdr2+/− mice; at the age of 2 months, the difference between mutant and control livers was even larger (left panels, Fig. 5B,C). Mbd1 staining was much weaker in the FVB strain versus the B6 strain, whereas the patterns of expression were similar and showed prominent localization in the hepatocyte nuclei (right panels, Fig. 5B,C). When we tested the expression levels of these proteins, Lipin-1 and Mbd1, in liver tumors from old Mdr2-KO mice of both strains, we found variable expression patterns for both proteins. They evidenced higher expression in some

tumors or nodules and lower expression in others in comparison with the noncancerous surrounding tissue (Supporting Fig. 5). Although genome-scale gene expression profiling

did not reveal aberrant expression of the Mat1a www.selleckchem.com/products/dinaciclib-sch727965.html transcript in the livers of Mdr2-KO mice of both strains, we compared Mat1a protein levels in total liver extracts from mutant and control mice at the age of 3 months. We found a significant decrease in Mat1a at the protein level in livers of Mdr2-KO/FVB mice but not in livers of Mdr2-KO/B6 mice (Fig. 6A,B). Both WT (Fig. 6A) and Mdr2+/− males (not shown) of the RAD001 ic50 FVB and B6 strains were used as controls, and they showed similar expression of Mat1a. Because of the paradoxical anti-inflammatory and protumorigenic effects of Gal-1 and its significant up-regulation in Mdr2-KO/FVB mice, we further validated the functional relevance of this endogenous lectin in both strains. MCE公司 We observed up-regulation of the Lgals1 transcript encoding the Gal-1 protein in the livers of 3-month-old Mdr2-KO/B6 mice by RT-PCR (Fig. 4C). We confirmed the increased expression of the Gal-1 protein in both mutant

Mdr2-KO livers (Figs. 5D and 6C,D) starting from the first month of age. The Gal-1 protein was localized mainly in the cytoplasm of nonhepatocyte cells, including immune cells and cholangiocytes, and rarely in the cytoplasm of hepatocytes. Remarkably, in the bile ducts, Gal-1 was detected only in proliferating cholangiocytes (ductular reaction). Expression of the Gal-1 protein increased between 1 and 3 months of age in both Mdr2-KO strains. When the strains were compared, the level of Gal-1 was higher in Mdr2-KO/B6 livers at the age of 1 month, but the levels were similar in the two strains at the age of 3 months (Fig. 6D). Previous studies showed that the administration of exogenous recombinant Gal-1 protected mice against autoimmune hepatitis induced by ConA.14 Here we demonstrate that B6 Gal-1–KO mice had an increased sensitivity to ConA-induced hepatitis (Fig. 7A,B), and this highlights the relevance of endogenous Gal-1. Reflecting hepatocyte injury, ALT blood levels were significantly higher in the B6 Gal-1–KO mice versus the control WT mice at every time tested (Fig.