Fig. 6

Chi3l1 KO mice have reduced pulmonary metastasis with increased IFNγ-producing CD4 and CD8 T cells in the lung. a Representative lung image from B16F10 melanoma injected WT and Chi3l1 KO mice. Scale bar, 2 mm b Number of pleural colonies in each lung was counted. Data are mean ± SEM of three sets of independent experiments and each dot in graphs represent an individual mouse. c Relative total tumor area in the lung was measured by Image J software 1.48 v. d Histology of lung sections by H&E staining, and infiltrated tumor region was measured by Image J software 1.48 v. Scale bar, 200 m. e, f IFNγ producing CD4 T cells, Foxp3 + regulatory T cells, IFNγ producing CD8 T cells, and Granzyme B expression in IFNγ⁺ CD8 T cells in the lung was analyzed by intracellular cytokine staining. % of IFNγ⁺, % of Foxp3⁺, and MFI of Granzyme B was represented as scattered graph. g, h IFNγ producing NK cells IFNγ producing non-lymphocytic population, and Granzyme B expression in IFNγ⁺ NK cells and non-lymphocytic population in the lung was analyzed. % of IFNγ⁺, and MFI of Granzyme B was represented as scattered graph. i mRNA expression of genes related to cytotoxicity and Th1 effector functions was analyzed by quantitative RT-PCR. Each gene expression level was normalized to β-actin. j Cytotoxicity of WT and Chi3l1 KO CD8 T cells against B16F10 target cells at indicated E:T ratios. k NK cell activity was represented as tumor killing activity of WT and Chi3l1 KO NK cells against to B16F10 target cells at indicated E:T ratios. Data are mean ± SD of three sets of independent experiments and each dot in graphs represent an individual mouse. n.s., not significant; *p < 0.05, **p < 0.01, ***p < 0.001 (two-tailed Student’s t-test)