Fig. 2

Immuno-FISH and CD8+ T-cell infiltrate in primary vs. metastatic samples. a iFISH images of one area of the primary tumor and brain and liver metastases of case 15. Scale bar 100  mm. b Dot plot depicting relative frequencies of the four different cell types in primary and metastatic samples. Error bars, S.E.M. c Immunofluorescence analysis of CD8, GranzymeB (GZMB), and pSTAT3. Images are a montage of nine fields captured from one area of the tissue. Scale bar 100  mm. d, e Graph depicting numbers of CD8⁺ T cells per montage (d) and fraction of GZMB⁺CD8⁺ T cells (e). Significance of the difference between primary and metastatic samples was calculated using the Wilcoxon rank-sum test. f Correlation analysis between the number of infiltrating CD8⁺ T cells and the relative frequencies of specific cell populations in primary and metastatic samples. g Correlation analysis between number of infiltrating CD8⁺ T cells and the Shannon index of diversity in all, primary and metastatic samples. Gray area, 95% confidence interval. Sample sizes were n  =  16 primary tumors and n  =  18 metastases