Article first published online: 25 APR 2012
Copyright © 2012 American Association for the Study of Liver Diseases
Volume 56, Issue 1, pages 322–331, July 2012
How to Cite
Singla, A., Moons, D. S., Snider, N. T., Wagenmaker, E. R., Jayasundera, V. B. and Omary, M. B. (2012), Oxidative stress, Nrf2 and keratin up-regulation associate with Mallory-Denk body formation in mouse erythropoietic protoporphyria. Hepatology, 56: 322–331. doi: 10.1002/hep.25664
Potential conflict of interest: Nothing to report.
Supported by NIH R01 DK52951 and the Department of Veterans Affairs (to M.B.O.); NIH Michigan Gastrointestinal Peptide Research Center P30 DK34933; and NIH F32 DK093202 (to A.S.).
- Issue published online: 3 JUL 2012
- Article first published online: 25 APR 2012
- Accepted manuscript online: 15 FEB 2012 04:12AM EST
- Manuscript Accepted: 27 JAN 2012
- Manuscript Received: 8 SEP 2011
Additional Supporting Information may be found in the online version of this article.
|HEP_25664_sm_SuppFig1.tif||307K||Fig.S1: Serum and biochemical analysis of livers of 13-week old mice. (A) Serum from 13-week old mice of the indicated genotypes were used to measure ALT, ALP and BA levels (N=3, *ast;p<0.05 compared to wt/wt). (B) RNA isolated from livers of 13-week old mice was used to quantify K8 and K18 mRNA levels. The ratio of K8 to K18 mRNA is 1.25 in fch/fch mice. *ast;p<0.05 compared to wt/wt. (C) Top panel (a) Coomassie stain for the HSEs (b) Immunoblot analysis of HSEs for K8 and ubiquitin. Ub crosslinks that appear at the top of the gel are highlighted by brackets. (c) total lysates were used to examine TG2 levels (β-tubulin is shown as a loading control).|
|HEP_25664_sm_SuppFig2.tif||3896K||Fig.S2: HE staining of 13-week old liver sections. Liver sections from 13-week old mice were HE stained to examine the MDBs. No MDBs were noted in the 13-week old fch/fch mice. Scale bar = 50 µm|
|HEP_25664_sm_SuppFig3.tif||9601K||Fig.S3: Immunofluorescence staining of 13-week old mouse liver sections. Liver sections from 13-week old mice were double-stained with K8/K18 (red) and ubiquitin (green) antibodies. No MDBs (yellow dots) were noted in the 13-week old fch/fch mice. Scale bar = 20 µm|
|HEP_25664_sm_SuppFig4.tif||7653K||Fig.S4: Immunofluorescence staining of 20-week old mouse liver sections. Keratins (red), ubiquitin (green) and nuclei (blue) staining of liver sections from 20-week old mice. Scale bar = 20 µm|
|HEP_25664_sm_SuppFig5.tif||607K||Fig.S5: Oxidative stress is upstream of proteasomal inhibition. FVB animals were fed 0.1% DDC for 2, 5 and 10 days. (A) Quantification of Nrf2 mRNA expression in control and DDC-treated livers. N=3, *ast;p<0.05. (B) Immunoblot analysis of the nuclear extracts from control and DDC-treated livers using anti-Nrf2 antibody. Coomassie stain of the nuclear extracts serve as loading control. (C) 20S proteasomal activity was measured in the liver lysates. N=3, *ast;p < 0.05. (D) Mitochondrial protein oxidation was measured and relative intensity of bands was quantified using ImageJ software. (E) Nrf2 levels are increased in MG132-treated HepG2 cells.|
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