Figure 8
Zone 1-HSC cells participate in sinusoid capillarization after CCl4 treatment. (a) SMMHC-tdTomato reporter mice were injected with tamoxifen (tam) on 5 consecutive days to induce the expression of the fluorescent dye tdTomato under control of the SMMHC promoter (SMMHC-tdTomato). 45 days later, animals received 5 injections of CCl4; controls did not receive CCl4. Animals were sacrificed, and livers were then isolated, fixed and analyzed. (b) Staining for CD31 and laminin (both green) revealed expression restricted to large vessels in control livers (b1 and b3) and strongly increased expression after fibrosis induction (b2 and b4). (c) tdTomato-expression (red, except for c4) was detected in laminin (green)-enriched sinusoidal/tubelike structures (asterisks/arrowhead, respectively). To more clearly show tissue structure (i.e., sinusoidal lumina), laminin is indicated by a gradient from blue to red and tdTomato is shown in yellow in c4. (d) Magnification of tdTomato/CD31 (d1) and tdTomato/laminin (d2) co-staining shows close association in hepatic sinusoids. (e) Triple staining of a single sinusoid for tdTomato, CD31 and laminin shows close association of endothelial cells, basal lamina and zone-1 HSC. Surrounding hepatocytes can be identified by DAPI-stained nuclei. Single channels are shown in e2-4.
