Fig. 2. JG-98 treatment reduces infiltration of macrophages into tumors.

A. JG-98 suppresses macrophage infiltration. Tumors established with HeLa/MDR1 cells were collected from control and JG-98-treated animals on day 6 after the start of drug administration, sectioned and stained with F4/80 antibodies. Brown color (arrows) indicates macrophages. B. Quantification of infiltration of macrophages, endothelial cells and fibroblasts in tumors. The sections were stained with F4/80, anti-CD31 (endothelium) and SMA (fibroblasts) antibody. Quantification was done in 4 fields from 3 tumors each. Data shown are mean of pixels per field +/−SD. C. Treatment of macrophages with JG-98 affects genes that regulate migration. RNA was isolated from macrophages that were treated with 1μM JG-98 or left untreated, and gene expression was assessed by microarrays. Genes related to motility/migration among 100 top hits are shown. The figure presents data from three independent experiments. Color coding show False Discovery Rate (FDR), which is the estimated probability that a gene with a given normalized enrichment score represents a false positive finding. In other words, blue colors reflect statistically significant downregulation of each gene compared to red colors. D. Enrichment profile of the RHO signaling pathway according to Gene Set Enrichment Analysis. Enrichment plot reflects FDR in a gene set. For more detailed explanation see http://software.broadinstitute.org/gsea/doc/GSEAUserGuideTEXT.htm E. Effects of JG-98 on macrophage migration. Macropahges were treated with indicated concentrations of JG-98, and migration was measured by the “wound healing” and transwell assays as described in Materials and Methods. Data shown are means +/− SEM of triplicates.