FIGURE 7. Expression of CX₃CL1 in human kidney and liver is higher compared to that in intestine/colon and omentum.

Immunohistochemical staining with CX₃CL1-specific antibodies was used to determine CX₃CL1 expression and intracellular localization in normal human specimens of liver and kidney (A), intestine/colon, and omentum (B). Brown, CX₃CL1; blue, hematoxylin. Typical images of CX₃CL1-immunoreactive staining in specimens of liver, kidney, intestine/colon, and omentum are shown. Image of liver corresponds to the case C6 (Supplementary Table 3), image of kidney corresponds to the case A4 (Supplementary Table 4), image of intestine/colon corresponds to the case C06 (Supplementary Table 1) and image of normal human specimen of epiploon (greater omentum) belongs to a 21 year old female. Brown, CX₃CL1; blue, hematoxylin. Images were generated using Aperio ScanScope; bar, 100 micron. (C) Comparison of the CX₃CL1 immunoscores among the tested tissues. The scoring was performed for the entire tissue, including all cell types present (of note, specimens of intestine/colon contained stroma underlying mesothelium, consistent with the microenvironment surrounding ovarian cancer cells anchoring in submesothelial stroma). The normal liver TMA contained 9 female and 15 male specimens; the average staining in female and male samples was 2.6 and 2.4, respectively (Supplementary Table 3). The normal kidney TMA contained 24 female and 56 male specimens; the average staining score in female and male samples was 2.1 and 2.3, respectively (Supplementary Table 4). Distal tubules displayed stronger immunoreactivity against CX₃CL1 antibodies compared to the proximal tubules. The visceral peritoneum TMA contained both stomach and intestine/colon specimens. Among those there were 13 female and 14 male intestine/colon specimens with average staining score of 1.2 and 1.0, respectively. Immunoscores of CX₃CL1 expression in female specimens of normal human liver, kidney, and intestine/colon were averaged and plotted. The data were analysed using both Mann-Whitney U test and Students’ t-test; * p<0.05.