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Supplementary material for this article can be found on the H EPATOLOGY website ( http://interscience.wiley.com/jpages/0270-9139/suppmat/index.html ).

FilenameFormatSizeDescription
fig1.tif9873KImmunolocalization of ECM components in adult mouse liver. (A) Type I collagen. (B) Type III collagen. (C) Type IV collagen. (D) Fibronectin. (E) Laminin. (F) Nidogen. (G) HSPG. Periportal connective tissue is positive for types I and III collagen (A, B). Staining of laminin and nidogen in sinusoids (E, F) is not strong as compared with that of collagen IV and HSPG (C, G). Fibronectin is localized around blood vessels, including sinusoids (D). Arrows indicate intrahepatic bile ducts. HV, hepatic vein; PV, portal vein. Bar indicates 50 μm.
fig2.tif10481KImmunolocalization of integrin subunits and PECAM-1 in adult mouse liver. (A) α1 integrin subunit. (B) α3 integrin subunit. (C) α5 integrin subunit. (D) α6 integrin subunit. (E) β1 integrin subunit. (F, G) β4 integrin subunit. (H) PECAM-1. Arrows indicate intrahepatic bile ducts, epithelial cells of which are positive for á3, á5,á6 and b4 integrin subunits (B, C, D, G), but negative for theá1 integrin subunit (A). α3 integrin signals are detected in periportal connective tissue (asterisk), but not in hepatic veins (B). β1 integrin signals are seen in sinusoids (E). β4 integrin subunits are expressed in endothelial cells of portal vein, but not in hepatic vein (asterisks, F, G). PECAM-1 expression is seen strongly in endothelial cells of the portal vein and weakly in those of sinusoids (H). HV, hepatic vein; PV, portal vein. Bars indicate 50 μm.
fig3.tif23866KDouble immunofluorescent analysis of nidogen (A, D, G, J, M) and cytokeratins (B), desmin (E), type III collagen (H), type IV collagen (K) or laminin (N) in 14.5-day fetal mouse liver. (C, F, I, L, O) Double exposure of A and B, D and E, G and H, J and K, and M and N. Periportal, cytokeratin-positive hepatoblasts are strongly immunostained for nidogen on the basal side (arrowheads, A-C). Nidogen deposition (asterisks) does not always colocalize with desmin-positi hepatic stellate cells (arrowheads, E, F) or type III collagen (arrowheads, H, I). Nidogen deposition (asterisks) is similar to localization of type IV collagen (arrowheads, K, L), but colacalizes with laminin (arrowheads, N, O). Arrows indicate basal lamina of periportal biliary epithelial cells (M-O). PV, portal vein. Bar indicates 50 μm.
fig4.tif4784KExpression of WGA-binding sites in 17.5-day (A) and 1-week-old livers (B). Fluorescent WGA strongly stains vascular structures in fetal and postnatal livers (arrows). HV, hepatic vein; PV, portal vein. Bar indicates 50 μm.
fig5.tif4159KImmunolocalization of laminin (A), and α3 (B) and α5 (C) integrin subunits in extrahepatic bile ducts. (A) 10.5-day liver. (B, C) 13.5-day liver. Epithelial cells of the extrahepatic bile duct are lined with laminin immunostaining (A). They are also a3 and a5 integrin subunit-positive while hepatoblasts are negative (B, C). Arrowheads indicate prospective hepatic duct regions that do not express these integrin subunits. Free cells are α3 integrin subunit-positive (asterisk, B). The liver region shows high positivity of α5 integrin subunits, which does not clearl delineate intrahepatic structures (C). ED, extrahepatic bile duct; HD, hepatic diverticulum; LP, liver parenchyma; PV, portal vein. Bar indicates 50 μm.

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