Fig. 4
Oncogenic role of KH-type splicing regulatory protein (KSRP) in vivo. A Dissemination of human clear cell renal cell carcinoma (ccRCC) cells in zebrafish embryos. Caki-1 cells were implanted into 48 h post-fertilization zebrafish embryos. Tumor cell dissemination was detected at day 4 post-injection. White arrows indicate the primary tumor and blue arrowheads indicate disseminated tumor foci. Frequencies of fish showing trunk and end-tail dissemination were monitored and counted every day (right panel). shCtrl: n = 12; shKSRP: n = 14. 4hpi, 4 h post-injection; 4dpi, 4 days post-injection. B Male NOD/SCID mice were orthotopically injected with luciferase-tagged and KSRP-knockdown (shKSRP), neural precursor cell-expressed developmentally downregulated 4 like (NEDD4L)-knockdown (shNEDD4L), or double-knockdown (shKSRP/shNEDD4L) Caki-1 cells. Whole-body bioluminescence imaging was conducted at the indicated time points after injecting cells into mice. C Quantitative analysis of Xenogen imaging signal intensity (photons/s/cm2/sr) every week. *p < 0.05, **p < 0.01, compared to the control group. ###p < 0.001 compared to the KSRP-knockdown only group. D Gross appearance of orthotopic tumors (indicated by the yellow dashed line) in a kidney. E Upper panel, Representative ex vivo bioluminescence imaging of metastatic sites (pancreas, liver, and lungs) at the end of this in vivo study. Lower panel, Signal intensities of metastatic organs were imaged with bioluminescence at the end of the study, with the mean signal for each group indicated. *p < 0.05, **p < 0.01, ***p < 0.001. F Caki-1 xenografts infected with the indicated shRNAs were isolated to detect expressions of vimentin and E-cadherin by IHC staining. Original magnification, 400 ×