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Figure 3—figure supplement 2. Cardiac lymphatic vessels extension is correlated with extent of the coronary arterial tree.

Whole-mount confocal imaging of adult transgenic zebrafish hearts expressing the pan-endothelial fli1a:GFP (green: A-L), Arterially and endocardially enriched endothelial marker kdrl:mTurquoise (red: A-L), lymphatic expressed flt4:mCitrine (blue: A-L). Estimated percentage coverage of surface fli1a:GFP expression within coronary vessel - as opposed to isolated cells or strings of such cells (green arrowheads) or non-lumenized conglomerates of cells (white arrowheads) - is listed for each image (Fli1a*) along with surface kdrl:mTurquoise and flt4:mCitrine coverage. Normally patterned coronary vessels show higher expression of kdrl:mTurquoise and are flanked by flt4:mCitrine-positivevessels on both sides of the heart (A and B). cxcr4a mutants show consistent abnormality in coronary vessel formation (C-L, range of mutant phenotypes including outliers). The degree of identifiably formed vessels varies in older mutants, but the spatial patterning of such vasculature over ventricle and the lack of kdrl:mTurquoise expression suggest no coronary artery tree structure. The extent of such coronary structure appears to be correlated with lymphatic extension down the ventricle, and is also largely absent in mutants. (M) Quantification of expression of surface kdrl:mTurquoise and flt4:mCitrine on individual sides of the heart (extension of arterial/lymphatic vessels is often more prominent on one or other side in wild type transgenic zebrafish, R2 = 0.36; p<0.0001). Scale bars 200 μm.

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