Endothelial-specific expression of grb2b is sufficient for normal lymphatic development. a In situ hybridization of grb2b shows expression in the majority of tissues in zebrafish embryos at 32hpf. b Rescue construct containing a 5xUAS element upstream of the grb2b cDNA which was fused with an RFP cassette via a P2A self-cleaving peptide. c?d? Confocal projections of transgenic flt4:Gal4; UAS:GFP; UAS:grb2b-P2A-RFP embryos from a tabula rasa in-cross at 5dpf. A tabula rasa wild-type embryo is shown in c?c? and a homozygous mutant in d?d?. e, f Quantification of trunk segments containing TD (scored over the length of 10 somites) in 5dpf embryos from a tabula rasa+/?; flt4:Gal4; UAS:GFP; grb2b-P2A-RFP in-cross. e GFP+ RFP? embryos (i.e., not containing the rescue construct) served as a control. wt: n?=?17; het: n?=?32; mut: n?=?18. f The mutant phenotype is rescued in embryos expressing the rescue construct (GFP+ and RFP+). wt: n?=?29; het: n?=?60; mut: n?=?19. TD thoracic duct, wt wild type; het heterozygous; mut mutant for tabula rasa. Scale bars: 50 µm
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