Higher magnification of MCs infiltrating the blastema and stroma is shown in Figure 2M. Therefore, both adaptive and innate immune cells were present in tumors Tanespimycin cost at a much higher frequency than in normal kidneys. Comparison of the various infiltrative inflammatory immune cells in tumors showed that the degree of TAM infiltration was significantly higher than the degree of infiltration by the other cells (Figure 3). Infiltration pattern of various inflammatory immune cells in different parts of the tumor was summarized in Table W2. Positive immunoreactivity for the COX-2 protein was observed in all tumors assessed relative to normal kidney (Figure 4, A–C). In most tumors,

weak to moderate cytoplasmic COX-2 expression was observed in blastemal and epithelial components and very intense nuclear staining was observed in the tumor stroma ( Figure 4C), although some of the tumors also showed intense cytoplasmic expression in blastemal and epithelial cells (not shown). Normal kidney samples showed weak to moderate

staining in the cytoplasm of some tubular epithelial cells ( Figure 4A) and very weak or no staining in renal interstitial cells or glomeruli. The sum density of COX-2 expression was significantly higher (about five times) in tumors than in normal kidneys ( Figure 4D). Although very little HIF-1 expression was noted in normal kidney slides (Figure 4E), seven of the seven tumors evaluated had cytoplasmic granular staining and membranous Fulvestrant purchase expression in blastemal and epithelial cells ( Figure 4F), with very prominent nuclear localization of HIF-1 protein expression

in the tumor stroma ( Figure 4G). The density of HIF-1 expression in tumors was significantly higher than that in control kidneys ( Figure 4H). The stromal expression of HIF-1 was similar to the COX-2 expression pattern ( Figure 4, C and G). Cytoplasmic expression of p-ERK1/2 in normal kidney was negligible (Figure 4I). In contrast, prominent nuclear p-ERK1/2 staining was observed in 10 of the 14 tumors analyzed. Although some expression in blastemal cells ( Figure 4J) was observed, p-ERK1/2 expression was primarily localized to tumor stroma ( Figure 4, GBA3 K and L). No p-ERK1/2 expression was observed in the epithelial component of tumors (not shown). Expression of p-ERK1/2 was significantly higher in tumors than in control kidneys ( Figure 4L). The stromal expression pattern of p-ERK1/2 was similar to those of COX-2, HIF-1, and VEGF. p-Stat3 expression was predominantly confined to the nucleus, with almost undetectable cytoplasmic staining in 10 of 13 the WT evaluated (Figure 4, M–O). No p-Stat3 expression was detected in three of the tumors. Almost all p-Stat3 expression was in blastema ( Figure 4N) or stroma ( Figure 4O). Very little or no p-Stat3 expression was observed in the epithelial component of the tumors (not shown). p-Stat3 expression was significantly higher in tumors than in normal kidney ( Figure 4P).