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Fig. 5 NAT10 facilitates tumor metastasis in vivo. (a) Three-dimensional overview of metastatic phenotype of the cancer cells at 1 dpi. Left, anterior. Up, dorsal. Right, lateral. Green, solid cancer cells, Red, transparent vessels. Imaging was acquired using Olympus confocal laser scanning microscope FV3000 at 40x magnification. Olympus CellSens software was used for multi-view fusion and deconvolution. Imaris 10.1 was applied for imaging stitching and 3D rendering. Intersegmental vessel (ISV), dorsal longitudinal anastomotic vessel (DLV), caudal artery (CA), caudal vein (CV). Scale bars: 20 μm. (b) BGC-823 cell expressing GFP and arresting within the tail vessel lumen at 1 dpi. Right panel shows quantification of distance from tumor cell to vessel wall for oeVector and oeNAT10 cells. Scale bars: 20 μm. (c) High-magnification confocal images of oeVector and oeNAT10 cells labeled with GFP that have arrested in the ISV lumen. Right panel shows quantification of distance from tumor cell to vessel wall for oeVector and oeNAT10 cells. Scale bars: 10 μm. (d) High-magnification images to show single cell behaviors. Single cells were extravasated from ISV, DLV, CA and CV. Right panel shows 3D rendering of intra- and extravascular cells. Up, extravasating tumor cells (green). Down, Circulating tumor cells (yellow). Scale bars: 20 μm. (e) Confocal image of GFP-expressing oeVector and oeNAT10 cells arrested in the ISV lumen and either inside or outside (extravasated) the vessel lumen. Image was taken 1, 2 days post injection (dpi) with a 10 objective. Right panel shows the percentage of extravasated tumor cells quantified 2 dpi for variants of BGC-823 cell line used in the manuscript: oeVector and oeNAT10. Scale bars: 10 μm.