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Fig. S3
Validation of the Tg(UAS:vegfc), Tg(shh:ccbe1) and Tg(shh:vegfc) transgenic line. (A) The Tg(4xUAS:vegfc) line, was crossed to the Tg(hsp70l:Gal4)(Scheer et al, 2001) to provide inducible ubiquitous expression of vegfc after heat-shock, or to the Tg(s1173:Gal4)(Scott & Baier, 2009) to provide specific expression in the tectal cell bodies, hindbrain, strong ocular muscles, and eye: bipolar cells. Schematic representation of the transgenic constructs. (B) In situ hybridization (as described (Schulte-Merker, 2002)) at 24hpf in a clutch of incrossed Tg(hsp70l:Gal4;4xUAS:vegfc) embryos. n=32/83 embryos displayed ectopic vegfc expression with an expected frequency of 50%. In a clutch of incrossed Tg(s1173:Gal4;4x- UAS:vegfc), 22/68 embryos displayed ectopic vegfc expression with an expected frequency of 50%. (C) Schematic representation of the shh:ccbe1 and shh:vegfc contructs. We generated these constructs using the MiniTol2 vector(Balciunas et al, 2006) to specifically overexpress either of these genes in the FP using the shh promoter and a FP specific enhancer. The effective expression of these genes can be monitored due to the integration of an IRES tagRFP or IRES mturquoise reporter. (D) Confocal projections in the Tg(fli1a:EGFP) background. At 56hpf in Tg(shh:ccbe1) embryos, RFP fluorescence is detected in the floorplate (FP).