Additional Supporting Information may be found in the online version of this article.

HEP_23998_sm_suppfig1.tif6313KSupplemental Figure 1. MCD diet-associated NASH is associated with upregulation of OPN expression. Mice were fed control chow or the MCD diet for 8 weeks, and then sacrificed to assess liver histology, serum ALT and hepatic OPN gene expression. (A) Representative H&E liver section (400X) and (B) serum ALT levels in WT mice. ALT was expressed as IU/L, and mean ± SEM. (C) Sirius red staining after MCD diet (200X) (D) Hepatic hydroxyproline content in both WT and Ptc+/- mice (E) OPN mRNA in WT mice. Results are expressed as fold change relative to control-chow fed mice; mean ± SEM. *P<0.05 vs. control-chow fed mice
HEP_23998_sm_suppfig2.tif3585KSupplemental Figure 2. Chow-fed WT and Ptc+/- mice contain similar amounts of OPN protein. WT and Ptc+/- mice were fed control chow or the MCD diet for 8 weeks, and then analyzed for OPN gene and protein expression. (A) Representative OPN immunostaining in chow-fed WT mice (200X) (B) Representative OPN immunostaining in chow-fed Ptc+/- mice (200X)
HEP_23998_sm_suppfig3.tif579KSupplemental Figure 3. OPN is Hedgehog-regulated and is upregulated during human HSC activation. Primary human HSC (ph HSC) were isolated and maintained till culture-activated to myofibroblasts (MF-HSC). Cells were then harvested for RNA and analyzed by QRTPCR. In a separate experiment, Day 4 ph HSC were treated with Cyclopamine (Cyc) or Tomatidine (Tom) for 24 hrs and OPN gene expression analyzed. (A) OPN mRNA in quiescent HSC, activated HSC (MF-HSC) and in the human HSC line, LX2. (B) OPN mRNA in Tom vs Cyc treated ph HSC. Results were expressed as fold change relative to Quiescent or Tomatidine-treated controls; mean ± SEM. *P<0.05 vs. controls.
HEP_23998_sm_suppfig4.tif7KSupplemental Figure 4. Liver is enriched with OPN in patients with ALD and NASH-cirrhosis. Total protein were harvested from explanted livers of individuals with ALD- and NASH-cirrhosis and analyzed by Western blot. Protein densitometry for OPN (n=3/group); results expressed as fold change relative to normal donor livers.
HEP_23998_sm_suppInfo.doc44KSupporting Information
HEP_23998_sm_supptables.doc55KSupporting Tables

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