In vivo invasion and dissemination of LEC-primed melanoma cells are dependent on MMP14 and Notch3.

(a) Time line of zebrafish xenograft experiments. (b) Intravital fluorescence microscopy images of six dpf zebrafish embryos taken four days post injection (4 dpi). Fluorescence in GFP channel is shown. Scale bar, 500 µm. Inset shows magnification of the primary tumor. Tumor cells invading outside pericardial space are marked with an arrow, invading cells in pericardial cavity with a triangle and unspecific fluorescence in eye and yolk sac with an asterisk (*). The outline of pericardial cavity is depicted with dashed line. (c) Quantification of area of primary tumors at 4 dpi. siCtrl, n = 52; siCtrl*, n = 74; siMMP14*, n = 61; siNotch3*, n = 46. (d) Quantification of circularity of primary tumors at 4 dpi. siCTRL, n = 52; siCtrl*, n = 74; siMMP14*, n = 61; siNotch3*, n = 46. (e) Quantification of melanoma cells invaded outside pericardial cavity. siCtrl, n = 43; siCtrl*, n = 55; siMMP14*, n = 44; siNotch3*, n = 37. (c–e) Non-parametric Kruskal-Wallis test with Dunn´s multiple comparison test was used, and in addition to individual data points, the median and interquartile range has been plotted. n.s., non significant (p>0.05); **p<0.01; ***p<0.001.