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. 2014 Nov 25;11:54. doi: 10.1186/1743-7075-11-54

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© Ito et al.; licensee BioMed Central Ltd. 2014

This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

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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 4 i. 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 5 a-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 5 g 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).