1C). The PHB2 protein level was also reduced, but to a lesser degree, to 30% to 40% of controls in the liver and hepatocytes (Fig. 1C). From the time of birth, there is variability in the weight and health of the KO mice. 15% of the pups (115/768) died before weaning (3 weeks old). Although most were not genotyped, of the ones that died before weaning and were examined, all were liver-specific KOs. KOs that survived past 3 weeks weighed less than WT control littermates and

this difference persisted up to 14 weeks of age (Supporting Figs. 2 and 3). The relative liver to body weight was higher in the KO mice (Table 1). At 3 weeks of age, many KO mice appeared ill, and liver injury is biochemically evident (Table 1). Liver injury is confirmed histologically by marked necrosis Olaparib price and inflammation seen throughout the liver Volasertib (Fig. 2B,C). There is also bile duct metaplasia (Fig. 2D), anisocytosis of hepatic nuclei (Fig. 2E), and positive staining for OV-6, an oval cell marker (Fig. 3B), and glutathione S-transferase Pi (GSTP) (Fig. 3D), a preneoplastic marker in the 3-week-old KO liver. Mat1a KO mice have higher hepatic triglyceride levels11

and develop steatohepatitis.12 This prompted us to measure lipid levels in the liver-specific Phb1 KO mice. Liver-specific Phb1 KO mice have elevated plasma cholesterol levels, but their hepatic cholesterol levels and both plasma and hepatic triglyceride levels were unchanged from WT controls (Table 1). As the mice grew older, by 14 weeks hepatic nodules can be seen in some liver sections but not

Dapagliflozin on gross examination (Fig. 2F). By 38 weeks, many KO livers stain positive for alpha-fetoprotein (AFP) (Fig. 3F). Because PHB1 is a mitochondrial chaperone protein, we examined mitochondrial morphology by EM. Supporting Fig. 4A,B shows that mitochondria in the 3-week-old KO liver appear swollen and many have no discernible cristae. Positive 4-hydroxynonenal (4-HNE) staining from increased lipid peroxidation in the KO liver, as compared to WT control liver (Supporting Fig. 4C,D), is consistent with impaired mitochondrial function. As the KO mice grew older, there was progressive apoptosis, as shown by activated caspase-3 staining (Fig. 4, top row), persistent proliferation as indicated by proliferating cellular nuclear antigen (PCNA) staining (Fig. 4, middle row), and progressive fibrosis on reticulin staining (Fig. 4, bottom row). Based on histologic examination, no frank cancer was noted in eight KO mice on a normal diet by 14 weeks. However, by 20 weeks, all mice have multiple liver nodules on gross examination of the liver (Fig. 5B); between the ages of 35 and 46 weeks 38% (5/13 mice; 1/5 male, and 4/8 female) have multifocal HCC (Fig. 5C,D). Because Phb1 KO mice develop HCC, we next compared PHB1 protein expression in normal primary human hepatocytes to that of human HCC cell lines Huh-7 and HepG2.