1. Localization of the Wilson's disease protein in human liver
Mark Schaefer, Han Roelofsen, Henk Wolters, Walter J. Hofmann, Michael Müller, Folkert Kuipers, Wolfgang Stremmel, Roel J. Vonk 
Gastroenterology 
Volume 117, Issue 6, Pages (December 1999)
DOI: /S (99)70288-X
Copyright © 1999 American Gastroenterological Association Terms and Conditions

2. Fig. 1
Immunoblot analysis of the WND protein in normal human liver. Immunoblot of enriched cLPM and blLPM incubated sequentially with WND antibody (WD, lanes 1 and 2), P-glycoprotein antibody (pgp, lanes 3 and 4), and NTCP (lanes 5 and 6).
Gastroenterology  , DOI: ( /S (99)70288-X)
Copyright © 1999 American Gastroenterological Association Terms and Conditions

3. Fig. 2
Immunoblot analysis of WND protein and AP-1–reactive material (trans-Golgi marker) in cLPM (lanes 3, 5, and 7) and blLPM (lanes 4, 6, and 8) fractions isolated from 2 separate human control livers (liver 1 and liver 2) and a liver from a subject with WND. A HepG2 cell lysate is shown as a reference in lane 1. Lane 2 contains the molecular mass markers. Note that in these preparations multiple bands are found for the WND protein, which probably reflects protein breakdown during storage of the livers or during the isolation procedure.
Gastroenterology  , DOI: ( /S (99)70288-X)
Copyright © 1999 American Gastroenterological Association Terms and Conditions

4. Fig. 3
Immunoblot analysis of the WND protein in a liver explant of a patient with WND. Equal amounts of protein (25 μg) of membrane fractions enriched in canalicular (WD cLPM) and basolateral membranes (WD blLPM) were separated by sodium dodecyl sulfate–polyacrylamide gel electrophoresis and sequentially immunoblotted with the WND antibody (upper blot) and the P-glycoprotein C219 antibody (lower blot). HepG2 cell lysate was used as a positive control.
Gastroenterology  , DOI: ( /S (99)70288-X)
Copyright © 1999 American Gastroenterological Association Terms and Conditions

5. Fig. 4
Immunohistochemical detection of WND protein in normal human liver. Liver sections were incubated with the WND antibody followed by a secondary antibody and visualized by an alkaline phosphatase–dependent reaction. The distribution of the WND protein can be seen in A. Some hepatocytes show a canalicular distribution pattern, whereas others show a vesicular distribution throughout the cytosol. Portal field with branches of portal vein (v), hepatic artery (a), and bile duct (b) and sinusoid (s) can be seen in B. Arrows indicate the punctuate intracellular staining pattern. Arrowheads outline the concentrated linear staining pattern of the WND protein at selective borders of hepatocytes (original magnifications: A, 80×; B, 160×).
Gastroenterology  , DOI: ( /S (99)70288-X)
Copyright © 1999 American Gastroenterological Association Terms and Conditions

6. Fig. 5
Double-label confocal scanning laser microscopy of the WND protein in human liver. To study the pericanalicular localization of WND protein in detail, frozen sections of normal and WND-affected liver were doubly labeled with antibodies against P-gp (A and D) to mark the canalicular membrane or against the Golgi markers 58K (G) and AP-1 (J) as well as with antibodies against WND protein (B, E, H, and K). The merged images are shown in C, F, I, and L. In normal human liver, WND-positive vesicles are clearly visible on both sides of the canalicular membrane, with weak staining of the canalicular membrane itself (B). Colocalization of P-gp and WND protein is not evident from the merged image (C), because of the differences in staining intensity. A clear P-gp staining (D) without any WND signal can be found in WND-affected liver (E). Both markers for the Golgi apparatus show a predominantly pericanalicular localization (G and J). As is evident from the merged images, 58K partially overlaps with the WND protein, whereas AP-1 almost completely overlaps, indicating that the WND-containing pericanalicular vesicles are part of the TGN (bars = 10 μm).
Gastroenterology  , DOI: ( /S (99)70288-X)
Copyright © 1999 American Gastroenterological Association Terms and Conditions