Fig. 5
xCT depletion in cancer cells impairs metastasis formation and alters the metastatic niche. A Western blot analysis of xCT expression in WT 4T1 cells and the pool of 10 xCTKO clones (xCT-KO Pool). Vinculin is used as loading control. B Growth curves of tumors deriving from 10.000 4T1 (WT or KO pool) injected subcutaneously in BALB/c mice. C Tumor weight at sacrifice. D Left: representative slices of FFPE lungs from mice described in panel B, stained with H&E. Right: percentage of slice area occupied by metastases. E Left: representative slices of FFPE lungs (following i.v. injection of 10.000 4T1, either WT or KO pool) stained with H&E. Right: percentage of slice area occupied by metastases. Percentage of selected immune cell populations over total leukocytes (CD45+) F infiltrating the lungs or G isolated from peripheral blood of tumor-bearing mice described in panel B. H Left: experimental protocol used to assess alterations in the immune pre-metastatic niche. Right: Mean tumor growth curves of mice are shown for a better visualization. Percentage of I PMN-MDSC or J NK over total leukocytes (CD45+) infiltrating the lungs of tumor-bearing mice (described in panel H) at different stages of tumor growth and of healthy, unchallenged mice. Number of replicates: number of mice is reported in panels B and H; each dot represents a mouse. For flow cytometry data, each dot depicting a mouse is the result of a single technical replicate. Statistical analysis: unpaired t test. * p<0.05; ** p<0.01; *** p<0.001. Where not indicated, p value is not significant. Lines (panel H only) and histograms represent mean values. Error bars are shown only when n > 5, and represent SD