The clinical and histological analysis after LPS-stimulation includes measuring of ALT (A) and AST (B).

<p>The histological analysis was performed using H&E staining (C). Overview pictures at 0 h were taken in 100-fold magnification, detail microphotographs at 3 h and 6 h post LPS such as infiltrating immune cells are magnified 200-fold (* = p<0.05; *** = p<0.005).</p

For determination of the liver injury FFPE-section were stained for DNA-strand breaks using a TUNEL-assay.

<p>Tunel⁺-cells were counted. Counterstaining was perfomed using Haematoxylin (A). Pictures were taken in 200 fold magnification. Results displaying an increase of TUNEL⁺-cells in regard to mFPR1 and mFPR2-deficiency are shown as a Graph (B). (* = p<0.05; ** = p<0.01.)</p

To identify the infiltrating immune cells after LPS stimulation according to their surface markers, CD11b was used to detect infiltrating monocytes and macrophages in the liver (A).

<p>Pictures were taken in 200-fold magnification and the CD11b⁺-cells were counted. The results were displayed as a graph (B) indicating differences among WT, mFPR1^-/- and mFPR2^-/- (* = p<0.05; ** = p<0.01; # = p<0.0001).</p

To better understand mechanism underlying the infiltration of immune cells into the liver of mFPR-deficient mice the gene expression of the pro-inflammatory cytokines IL-6 (A), TNF-α (B), CXCL1 (C), TLR2 (D) and TLR4 (E) were analyzed by qPCR.

<p>Changes in gene expression were related to GAPDH as a housekeeping gene. (* = p<0.05; ** = p<0.01; # = p<0.0001.)</p

qPCR Primer.

<p>qPCR Primer.</p

The Expression of the three mFPRs mFPR1 (A), mFPR2 (B) and mFPR3 (C) was analysed by qPCR and the gene expression was related to GAPDH.

<p>For the analysis of inflammation in the BDL-model immune cells were stained for CD11b-positivity. Graphical score for the amount of CD11b⁺-cells in the liver displays an increase over time. The cells are displayed as numbers per view field (D).</p

The second subset of immune cells were analysed by Ly6G-staining.

<p>Mainly neutrophil granulocytes were identified as Ly6G⁺ (A). Pictures were taken in 200-fold magnification and the Ly6G⁺-cells were counted. The results were displayed as a graph (B) indicating differences among WT, mFPR1^-/- and mFPR2^-/- (* = p<0.05; ** = p<0.01; # = p<0.0001).</p

To investigate liver proliferation FFPE-sections were stained with the universal cell cycle marker Ki67.

<p>At 3⁺-nuclei were counted and analyzed as percentage of proliferative cells. Photomicrographs were taken at 200-fold and representative images are shown. Ki67⁺-nuclei are indicated by arrows (* = p<0.05; ** = p<0.01).</p