Fig. 1

vegfc is not essential for zebrafish early facial lymphatic development. (A-J) Confocal images of the facial lymphatics in lyve1 embryos at 3dpf (A-E) or trunk lymphatics at 5 dpf (F-J) in control MO (A,F), vegfc MO-1 (B,G), vegfc^hu5055 (C,H), sFLT4 mRNA (D,I), and ccbe1 MO (E,J). Loss of Vegfc prevents formation of the TD (red asterisks), but the LFL forms normally at 3 dpf (white arrowheads) in vegfc morphant or vegfc^hu5055 mutant embryos. (K) Quantitation of LFL length at 3 dpf. (L) Quantitation of TD formation at 5 dpf. (M-Q) Confocal images of the facial lymphatics in lyve1 embryos at 5 dpf in control MO (M), vegfc MO-1 (N), vegfc^hu5055 (O), sFLT4 mRNA (P), and ccbe1 MO (Q). Knockdown of Flt4, Ccbe1 or Vegfc inhibits the correct development of the facial lymphatic network. Asterisk indicates the PHS. (R) The percentage formation of different facial lymphatic vessels at 5dpf. For flt4^hu4602 and vegfc^hu5055 embryos, n=number of mutant embryos. CCV, common cardinal vein; PHS, primary head sinus; PCeV, posterior cerebral vein; LFL, lateral facial lymphatic; OLV, otolithic lymphatic vessel; MFL, medial facial lymphatic; LAA, lymphatic branchial arches. ***P<0.001, by a Mann–Whitney test versus control MO; error bars indicate s.d. Scale bars: 50 μm.