|ZFIN ID: ZDB-IMAGE-180817-3|
Vascular specific overexpression of Flt4 is not sufficient to rescue lymphangiogenesis defects in photoactivated etv2 knockdown embryos.
(A,B) The number of lymphatic prox1a:GFP cells is reduced at 30 hpf in embryos injected with etv2 Photomorph solution and photoactivated at 24 hpf (2.4±0.6, n=7) compared to control uninjected embryos (7.5±0.8, n=8, p<0.001, two-tailed Student’s test). Note that prox1a reporter expresses both RFP and GFP.
(C) Injection of fli1a:flt4-2a-mCherry results in a reduction of the number of prox1a:GFP+ cells at 30 hpf in wild type embryos (2.8±1.0, n=5, p=0.004).
(D) Co-injection of fli1a:flt4-2a-mCherry together with etv2 Photomorph solution followed by photoactivation at 24 hpf results in a reduced number of prox1a:GFP cells (3±0.4, n=6, p=0.48 compared with the injection of etv2 Photomorph alone, panel B). (A’,B’,C’,D’) are higher magnification images of (A,B,C,D).
(E) Quantification of prox1a:GFP cell number in the trunk region (across 9 somite length) in different groups. Each data point corresponds to the cell number in a single embryo. Mean±SEM values are shown. Arrowheads or blue pseudocolor indicate prox1a:GFP+ cells within the PCV.
(F,G) Stable transgenic fli1a:flt4-2a-mCherry larvae (G,G’) have defective thoracic duct development at 5 dpf compared to fli1a:GFP larvae (F,F’). Each experiment has been repeated a minimum of two times.
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