Figure 5

Co-localization of ALDH1A1 protein expression and stellate cell/fibroblast marker, vimentin in liver from VAA rats. Results for VAD rats were similar and therefore are not shown. Dual IHC with anti-ALDH1A1 antibody and anti-vimentin antibody was performed followed by methyl green counterstaining for detection of nuclei. Staining controls for vimentin were similar to those shown for ED1 in Figure 4i. Panels c and d show false-color images after ilastik® processing (see Methods) so that only pink (vimentin) signals are visible as black. Arrows illustrate some of the cells that co-stained with purple (ALDH1A1) and vimentin signals (Figure 5a-f). Figure 5e illustrates the intense ALDH1A1 staining around bile duct structures (black arrow) and its absence around the arterial smooth muscle region (white arrow), which is also apparent in other sections. Magnification x 400. Figure 5g shows the tissue area occupied by vimentin staining, analyzed by ilastik®, which was significantly higher in the liver of rats treated with LPS (n = 8 animals) compared to those not treated with LPS (n = 9 animals, P = 0.011).