1. A
ID scr sh cx cx cx6 B C
Supplementary Figure 4. The fractalkine axis does not regulate ascites formation. Survival (A) and ascites formation (B) in animals bearing tumors formed by i.p. injecting ID8, scr sh, cx2, cx3, and cx6 cells as well as PBS into abdomens of C57BL/6 mice. Mice injected with phosphate buffered saline (PBS) served as a control; animals in this group did not develop any pathologic conditions over the course of the experiments and were sacrificed at the time when the last experimental animal reached humane endpoints, i.e. 515 days after i.p. injection of PBS, reaching a total lifespan of 1.5 years. Kaplan-Meier plot was used to estimate survival. Median survival was found using Prizm software and plotted on the graph. Difference in survival between the experimental (cx2, cx3, cx6) and control groups (ID8, scr sh) was statistically analysed using Log-rank (Mantel-Cox) test and Gehan-Breslow-Wilcoxon test; p>0.05. Color coding for the control and experimental animal groups : blue – PBS, red – ID8, green – scr sh, purple – cx2, orange – cx3, black– cx6. Typical image of animals presenting with ascites is shown (B, right panel). (C) Expression of CX3CL1 in specimens of lymph node was detected using immunohistochemistry. Image of CX3CL1 immunostaining in normal lymph node of epiploon corresponds to the case D7 (Supplementary Table 2). Brown, CX3CL1; blue, hematoxylin. Images were generated using Aperio ScanScope. The right panel is a magnified image outlined in the left panel. Bar, 500 (left panel) and 100 (right panel) micron.