Fig. 5.
Activated fibrogenic response in Ncu-g1gt/gt liver. (A,B) Immunofluorescence image shows CD68- and F4/80-stained KCs (green). Liver sections were counterstained with DAPI (blue). Infiltrating leukocytes can be seen in Ncu-g1gt/gt liver section (white arrow). CD68-positive cells seem to congregate at the site of leukocyte infiltration. In Ncu-g1gt/gt liver sections, F4/80-positive cells appear swollen and hypertrophic. Scale bars: 50 μm. (C) qPCR analyses of the restorative (Cd81) and the pro-inflammatory (Cd86) macrophage markers show elevated expression of only Cd86 in Ncu-g1gt/gt liver. (D,E) Activated KCs produce pro-fibrogenic cytokines. qPCR analyses show elevated expression of genes characteristic for activated fibrogenesis (Tnf, Pdgfb, Tgfb1, Tgfbr2). (F) Presence of activated HSCs was indicated by increased expression of genes characteristic for activated HSCs (α-Sma, Igfbp3, Vim). (G) This was supported by α-Sma staining in liver sections from wild-type (WT) and Ncu-g1gt/gt mice. Note the positive labeling of α-Sma indicating activated HSCs (arrows) in affected areas. Liver sections were counterstained with hematoxylin. Scale bar: 50 μm. (H) Immunofluorescence imaging shows increased levels of myeloperoxidase at the protein level in Ncu-g1gt/gt liver (green). Liver sections were counterstained with DAPI (blue). Note the granular staining pattern of myeloperoxidase, clustering of myeloperoxidase-positive cells and increased levels of invading polymorphonuclear leukocytes with small nuclei. Scale bar: 50 μm. (I) qPCR analyses supported the elevated myeloperoxidase (Mpo) expression in Ncu-g1gt/gt liver. (J) The expression of S100 calcium binding protein A8 (S100a8) was also elevated in Ncu-g1gt/gt liver. (C-F,I,J) n=4, *P<0.05, **P<0.005, ***P<0.001. Values are expressed as mean±s.e.m.