Supplementary material for this article can be found on the H EPATOLOGY website ( ).

jws-hep.20889.fig1.TIF2591KRepresentative illustrations of desmin expression in HSC. (A) Rat liver 3 d after intrasplenic injection of 1×10 6rat hepatocytes. (B) Rat liver 3 d after intraportal injection of 2×10 7MAA particles. Note that injection of MAA particles produced extensive desmin response in HSC, particularly in ischemic areas, where phagocytes can be observed along side desmin-positive HSC. This was in agreement with the finding that ischemic events associated with mechanical occlusion of liver sinusoids was sufficient for HSC activation. However, in recipients of only 1×10 6hepatocytes, which did not reproduce ischemic injury in the liver, owing to the small number of cells injected, we still observed activation of desmin expression in HSC, including HSC in the immediate proximity of transplanted cells, as well as away from transplanted cells. This is seen in panel A, where DPPIV-positive transplanted cells (in red color, arrows) and adjacent desmin-positive HSC (arrowheads) are observed. Therefore, activation of HSC following cell transplantation was not simply due to ischemic injury in the liver. The data were obtained with desmin immunostaining alone (B) or in combination with DPPIV histochemistry (A). Tissues were lightly counterstained with hematoxylin. Orig. Mag., ×200.
jws-hep.20889.fig2.TIF2862KDesmin immunohistochemistry of the liver is shown in representative animals 3 d after transplantation of 2×10 7hepatocytes. (A) Control rat treated with intrasplenic injection of normal saline with cells, showing desmin-positive HSC mostly in zone 1 of the liver lobule. (B) Showing a rat treated with intrasplenic injection of NTG along with cell transplantation. Note presence of more desmin-positive HSC in this situation, as well as their distribution in additional areas of the liver lobule, besides zone 1. Pa = portal area; Cv = central vein. Orig. Mag., ×40.
jws-hep.20889.fig3.TIF4184KShowing the effect of gliotoxin on HSC and Kupffer cells in animals treated with CCl 4plus vehicle (A, C, E) or CCl 4plus gliotoxin (B, D, F) 24 h previously. Panel A shows large numbers of desmin-positive HSC, including in periportal areas (Pa), although more prominently in perivenous areas of the liver. (B) The numbers of desmin-positive HSC in CCl 4and gliotoxin treated rats decreased. Panels C and D show higher magnification views of the periportal areas indicated by (Pa*) in panels A and B to again indicate depletion of desmin-positive HSC in D. Panels E and F show carbon containing Kupffer cells throughout the liver lobule, without any differences in animals treated with CCl 4plus either vehicle alone or gliotoxin. Panel G shows replicate western blots for α-SMA or PDI. Lanes 1–4 are from a rat treated with CCl 4plus vehicle alone with 1:2, 1:5 and 1:10 dilutions of the liver lysate in lanes 2, 3 and 4, respectively. Lane 5 is from a recipient of CCl 4plus gliotoxin, and lane 6 is from an untreated normal control rat liver. Note decreased α-SMA expression in the gliotoxin-treated rat liver (lane 5 versus lane 1). However, this decrease was not observed in all experiments using animals treated with CCl 4plus gliotoxin. Orig. Mag., A and B, ×40; C-F, ×200.
jws-hep.20889.mtd-cit.doc44KExperimental Methods, Results, Supplementary References

Please note: Wiley Blackwell is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.