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Fig. 2 - Supplemental 4 Metastatic spreading following activation of KRASG12V and Ventx (KR+VX). (A) Schematic representation and confocal microscopy of zebrafish head showing the early malignant cells (H2B-mTFP+ blue cells; white arrowheads) in the brain. Green EOS fluorescence has been used to visualize the whole zebrafish head. (B) Short-range dispersal in the brain of the early progeny of the initial malignant cell (H2B-mTFP+ blue cells; red arrowheads). An asterisk (*) indicates the eye and a white arrow the otic vesicle. (C) Schematic representation of the zebrafish larva trunk (left, dotted square), where H2B-mTFP+ cells (white arrowheads in center panel) can localize in KR+VX larvae, both as isolated cells (as in the dotted square a, 44.5 µm of mean distance) and as small clusters (as in the dotted square ?, r=22.7 µm). (D) Schematic representation of zebrafish larva digestive tract (left, dotted square) and confocal microscopy showing H2b-mTFP+ cells (white arrowheads, center and right panel) localized in the gut. Green EOS fluorescence has been used to visualize the digestive tract. Red arrows indicate the bulb, the green arrows point to the mid-gut, the blue arrows the distal-gut, and the blue asterisk (*) indicates to the gut lumen. Scale bars and body axes (a: anterior; p: posterior; d: dorsal; v: ventral) are shown.