Fig. 9.

In vivo administration of dNP2-siChi3l1 complex inhibits pulmonary metastasis with enhanced Th1 and CTL effector molecules. a Experimental scheme of dNP2-siRNA complex treatment in pulmonary melanoma metastasis model. b Representative lung image of dNP2-siEGFP and dNP2-siChi3l1-treated mice. Scale bar, 2 mm c 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. d Relative total tumor area in the lung was measured by Image J software 1.48 v. e 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 f, g IFNγ producing CD4 and CD8 T cells, and Granzyme B expression level in IFNγ⁺ CD8 T cells in the lung was analyzed by intracellular staining. % of IFNγ⁺, and MFI of Granzyme B was represented as scattered graph. h Intracellular T-bet expression level in CD4⁺IFNγ⁺ and CD8⁺IFNγ⁺ population. MFI 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. 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)