These authors contributed equally to this work.
Liver-specific deletion of prohibitin 1 results in spontaneous liver injury, fibrosis, and hepatocellular carcinoma in mice†
Version of Record online: 1 OCT 2010
Copyright © 2010 American Association for the Study of Liver Diseases
Volume 52, Issue 6, pages 2096–2108, December 2010
How to Cite
Ko, K. S., Tomasi, M. L., Iglesias-Ara, A., French, B. A., French, S. W., Ramani, K., Lozano, J. J., Oh, P., He, L., Stiles, B. L., Li, T. W.H., Yang, H., Martínez-Chantar, M. L., Mato, J. M. and Lu, S. C. (2010), Liver-specific deletion of prohibitin 1 results in spontaneous liver injury, fibrosis, and hepatocellular carcinoma in mice. Hepatology, 52: 2096–2108. doi: 10.1002/hep.23919
Potential conflict of interest: Nothing to report.
- Issue online: 23 NOV 2010
- Version of Record online: 1 OCT 2010
- Accepted manuscript online: 20 AUG 2010 12:00AM EST
- Manuscript Accepted: 6 AUG 2010
- Manuscript Received: 27 APR 2010
- National Institutes of Health (NIH) grant. Grant Numbers: DK51719, AA8116, AT1576
- USC Research Center for Liver Diseases grant. Grant Numbers: P30DK48522, SAF 2008-04800, HEPADIP-EULSHM-CT-205, ETORTEK-2008
Additional Supporting Information may be found in the online version of this article.
|HEP_23919_sm_suppfig1.tif||204K||Supporting Fig. 1. Schematic procedure of creating liver-specific Phb1 KO mouse. A new Phb1 mouse genomic contig spanning 39,999 bp was designed and cloned, containing neomycin sequences flanked by FRT sites and Phb1 exon 2 flanked by loxP sites. The cloned vector was introduced in the mouse genome via ES cell transfection and selected by neomycin screening. The neomycin-resistant ES cells were injected into blastocysts, and the chimeric mice were bred with normal C57BL/6J mice. Neomycin sequence was then deleted by breeding with Flp-recombinase transgenic mice. After developing the Phb1loxP/loxP homogeneous mouse strain, they were bred with Alb-Cre+/+ mice to produce Phb1loxP/+ with Alb-Cre+/−. Breeding of Phb1loxP/+ with Alb-Cre+/− resulted in Phb1loxP/loxP with Alb-Cre+/+ or with Alb-Cre+/−. loxP, locus of X-over P1.|
|HEP_23919_sm_suppfig2.tif||85K||Supporting Fig. 2. Periodic BW changes in liver-specific Phb1 KO mice. The graph shows the difference of BW changes between WT control and liver-specific Phb1 KO male mice. Liver-specific Phb1 KO mice display lighter BW from their weaning till 14 weeks after parturition. Observation numbers in each time point are 2 to 19 in liver-specific Phb1 KO and 2 to 8 in WT control mice. The data are expressed as mean ± SEM; *P < 0.00006. BW, body weight; SEM, standard error.|
|HEP_23919_sm_suppfig3.tif||5862K||Supporting Fig. 3. Physical appearance of WT and liver-specific Phb1 KO mice at different ages.|
|HEP_23919_sm_suppfig4.tif||5944K||Supporting Fig. 4. Abnormal mitochondria and oxidative stress in the liver-specific Phb1 KO livers. Mitochondrial morphology on EM of the livers from 3-week-old WT control (A) and liver-specific Phb1 KO mice (B) (×12,500). Liver-specific Phb1 KO mouse showed swollen mitochondrial morphology. Some of the swollen mitochondria formed dense inclusions (arrows). (B) This indicates that the liver cell is undergoing early signs of necrosis; the mitochondrial cristae areas are largely gone. The degree of oxidative stress in the livers from (C) 3-week-old WT control (×400) and (D) liver-specific Phb1 KO (×400) is shown by 4-HNE staining (brown cytoplasmic staining), which shows increased staining in the KO hepatocytes. The 4-HNE–positive cells show early signs of necrosis.|
|HEP_23919_sm_suppfig5.tif||1741K||Supporting Fig. 5. Heat map showing hepatic differential gene expression from WT and liver-specific Phb1 KO mice.|
|HEP_23919_sm_suppfig6.tif||6046K||Supporting Fig. 6. PHB1 protein subcellular localization in hepatocytes. Red (upper left) shows the location of mitochondria. Green (upper right) represents the location of PHB1 protein. Blue (bottom left) exhibits the location of nuclei in the cells. Bottom right is the merged image of red, green, and blue. (A) Localization of PHB1 protein in hepatocytes of the WT mouse shows it is mainly colocalized with mitochondria but also in the nuclei. (B) Hepatocytes of liver-specific Phb1 KO mouse express less amount of PHB1 protein in both compartments. (C) Normal mouse hepatocyte cell line (AML12) expresses PHB1 protein in the same locations as mouse hepatocytes.|
Please note: Wiley Blackwell is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.