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Figure 3 Whole-mount confocal imaging of adult transgenic zebrafish hearts expressing arterial marker cxcr4a:mCitrine (green: A and B), and lymphatic endothelial markers lyve1:RFP (Red: A and C) and flt4:mCitrine (Blue: D-F), the pan-endothelial marker fli1a:GFP (green: D-F), and the arterially-enriched pan-endothelial marker kdrl:mTurquoise (red: D-F). (A-C, n = 8) cxcr4a:mCitrine expression is restricted to the coronary arteries and not detectable in lyve1:RFP-positive LECs. (D) Cardiac lymphatic vessels in control (cxcr4a+/+, n = 8) zebrafish. (E and F) In cxcr4a mutants (cxcr4a-/-, n = 12) that have abnormal and discontinuous coronary vasculature the cardiac lymphatic extension is stunted. (G) Graph showing significant reductions in vessel density, kdrl expression in blood vessels and flt4 expression on the surface of the cxcr4a-/- ventricle. There is comparable levels of fli1a-expressing cells on the surface of the ventricle between larger/older cxcr4a-/- and control zebrafish, but loss of the coronary structure. (H) Schematic representation of the regulation of cardiac lymphatic developmental guidance by coronary arteries. Scale bars 200 μm.