, Waltham, MA) and utilizing a label-free approach. Two independent replicate MS analyses were carried out per sample. Data are represented as the mean ± standard error of mean (SEM) and were analyzed for statistical significance using one-way analysis of variance, MK2206 followed by Newman-Keuls’

test as a post-hoc test. A P value of <0.05 was considered as significant. We have created a TG mouse in a B6/CBA background with hepatocyte-specific expression of human AEG-1 by using the mouse ALB promoter/enhancer element to drive AEG-1 expression. This particular strain of mouse was chosen because it is very sensitive to hepatocarcinogenesis induced by DEN.11 The human AEG-1 has a C-terminal HA-tag. The expression of AEG-1 in the liver of Alb/AEG-1 mice was confirmed by western blotting Inhibitor Library ic50 analysis

using anti-HA antibody (Ab) (Fig. 1A). Two founder lines were characterized, initially revealing no significant differences. We therefore pursued further characterization employing one founder line. Male WT and Alb/AEG-1 littermates were given a single IP injection of DEN (10 μg/g) at 14 days of age and were monitored every 4 weeks, starting at 20 weeks. At 28 weeks of age, only 2 of 11 WT animals showed a few very small nodules in the liver, whereas all of the 17 Alb/AEG-1 mice livers harbored numerous nodules of different sizes (arrows in Fig. 1B,C). There was a significant increase in liver-to-body-weight ratio in Alb/AEG-1 mice, when compared to that in WT (Fig. 1D). Histological analysis of the livers of WT mice showed

a few dysplatic, hyperchromatic nuclei (arrow in Fig. 2A), indicating that, with time, HCC would eventually develop. In Alb/AEG-1 mice, a marked increase in dysplastic, hyperchromatic nuclei was observed both in the nodules as well as in the adjacent healthy liver (arrows in Fig. 2B,C). The most striking feature was observed in the hepatic nodules of Alb/AEG-1 mice, showing profound steatotic phenotypes with large lipid droplets in the hepatocytes (Fig. 2C). A moderate level of steatosis was also observed MCE公司 in the adjacent healthy liver in Alb/AEG-1 mice. There was a significant increase in hepatic enzymes in the sera of Alb/AEG-1 mice versus the sera of WT mice (Supporting Fig. 1). At 32 weeks of age, the WT mice developed hepatic nodules; however, the nodules that developed in Alb/AEG-1 mice were markedly larger (Supporting Fig. 2). These findings indicate that AEG-1 significantly accelerated the hepatocarcinogenic process in DEN-treated animals. The WT and Alb/AEG-1 mice were followed for 1 year without any DEN treatment. Although AEG-1-induced steatosis was profoundly evident, overt nodular HCC did not develop at this time point.