Fig. 4. Blockade of metastasis by NCKAP1 knockdown in animal models.

A A liver metastasis animal model was developed via intrasplenic injection of shNCKAP1-transfected HCT116-Luc cells (5 × 10⁶ cells/50 μl), and splenectomy was performed after 5 min of circulation in BALB/c nude mice. Images of metastatic tumor growth were monitored by the IVIS® Spectrum at 0, 7, and 19 days. Mice with metastatic livers were sacrificed at the endpoint of 22 days. Quantitative results on the right: the graph shows the fold change of liver weight/body weight ratio, and data are reported as the mean ± SEM. One-way ANOVA and Bonferroni’s post hoc test. *p < 0.05 vs. Control. (Control n = 4, shNCKAP1 (1) n = 3, shNCKAP1 (2) n = 2). B Tumor tissues for orthotopic transplantation were prepared from shScramble or shNCKAP1-transfected HCT116-Luc cells (5 × 10⁶ cells/50 μl) tumor tissues grown subcutaneously in the BALB/c nude mice. The tumor growth was monitored by for 38 days. Livers were isolated on day 38, and images of liver metastasis were measured by the IVIS® Spectrum.