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

HEP_25758_sm_SuppFig1.tif6392KSupporting Information Figure 1. Immunohistochemistry of SOX9 and OPN in normal mouse liver. Consecutive 5 μm sections of normal liver in mouse stained for SOX9 and OPN (brown), counterstained with toludine blue. Detection only in the round nuclei (SOX9) and cytoplasm (OPN) of biliary epithelial cells. Size bar represents 50 μm
HEP_25758_sm_SuppFig2.tif685KSupporting Information Figure 2. Sox9 and Opn detection in activated HSCs. A, Example immunoblot showing increased Sox9 and Opn in activated (A) compared to quiescent (Q) rat HSCs (rHSCs). B, Quantification of Sox9 and Opn in quiescent and activated rHSCs detected by immunoblotting. Immunoblotting quantification was normalized to β-actin. **, p<0.01, †, p<0.005.
HEP_25758_sm_SuppFig3.tif159KSupporting Information Figure 3. PCR for an intergenic region of GAPDH (i.e. no Sox9 binding motif) as a negative control for the ChIP assay shown in Figure 4. Positive control is input (diluted 10-fold). No PCR product is seen following immunoprecipitation using anti-Sox9 (or negative control IgG) antibodies.
HEP_25758_sm_SuppFig4.tif2893KSupporting Information Figure 4. A and B, Example immunoblots showing decrease and increase in OPN and SOX9 following cyclopamine or SAG treatment respectively (relating to experiments in Fig. 6B and D). C, Example immunoblot for Fig. 6E). D, Control data for Fig. 6F to demonstrate that transient transfection of the SOX9 cDNA expression vector produces increased levels of SOX9 in the presence of cyclopamine (CYC) in LX2 cells. Quantification of SOX9 protein was normalized to β-actin. †, p<0.005 compared to empty vector (EV) / DMSO treated control.
HEP_25758_sm_SuppFig5.tif475KSupporting Information Figure 5. A and B, example immunoblots and verification of myc-tagged GLI2?N and GLI3A expression in cells following transfection. EV, empty vector control.
HEP_25758_sm_SuppInfo.doc35KSupporting Information
HEP_25758_sm_SuppTabs.doc67KSupporting Information Tables.

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