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Fig. 2 Lymphatic vessel expansion during edema recovery. (A) Confocal image of osmotically balanced Tg(mrc1a:egfp; kdrl:mcherry) larvae illustrating the normal vascular network at 9 dpf. (B) Lymphatic vessels are pseudo-colored in green from panel (A) (C) Magnified image of the thoracic duct in panel (B) (D) Confocal image of recovered edema-free Tg(mrc1a:egfp; kdrl:mcherry) larvae illustrating the vascular network following edema resolution. (E) Lymphatic vessels are pseudo-colored in green from panel D, highlighting the excess lateral lymphatics and dilated thoracic duct. (F) Magnified image of the thoracic duct in panel E. (G) Quantification of the thoracic duct surface area in controls (n = 14) and edema-resolved larvae (n = 19). (H) Quantification of the number of DLLV nuclei in controls (n = 7) and edema-free larvae (n = 5) in Tg(fli1a:nEGFP; lyve1b:dsRed) background. (I) Comparison of the number of LL segments in 9 somites between controls (n = 11) and edema-free larvae (n = 10). (J) Quantification of the number of intersegmental lymphatic vessels in controls (n = 7) and edema-free larvae (n = 5). (K) Confocal image of the facial lymphatics of osmotically balanced larvae at 9 dpf. (L) Lymphatic vessels are pseudo-colored in green from panel K. (M) Magnified image from panel L. (N) Confocal image of the facial lymphatics of recovered edema-free larvae at 9 dpf. (O) Lymphatic vessels are pseudo-colored in green from panel N. (P) Magnified image from panel O showing excessive lymphatic sprouts. (Q) Comparison of the total facial lymphatic area in controls (n = 7) and edema-free larvae (n = 5). (R) Number of facial LEC nuclei in controls (n = 5) and edema-free larvae (n = 5). Tg(fli1a:nEGFP; lyve1b:dsRed) was used for quantitation. Scale bar = 50 μm (A-L). *p < 0.05, **p < 0.01; ****p < 0.0001.