SoxF factors are required for notch1b-15 activity. (A) Confocal projection of stably transgenic WT tg(notch1b-15:GFP) (left) and tg(notch1b-15mutSOX-a/b:GFP) (right), where both zfSOX-a and zfSOX-b sites have been simultaneously mutated. Representative larvae from four independent founders are shown. Asterisks indicate the reduced GFP expression in the dorsal aorta. Arrowheads show the ectopic neuronal expression observed in tg(notch1b-15mutSOX-a/b:GFP). All panels show composite images from tile scan acquisition. (B,C) The intensity of GFP expression in the dorsal aorta and arterial intersomitic vessels of the stably transgenic tg(notch1b-15:GFP) and tg(notch1b-15mutSOX-a/b:GFP) at 2 dpf. Expression is normalised to GFP genomic copy number. Fish were pooled from three or four separate founders. Mean±s.e.m. tg(notch1b-15), n=41; tg(notch1b-15mutSOX-a/b:GFP), n=29. ****P<0.0001 (Mann–Whitney U-test). (D) Levels of GFP expression in tg(notch1b-15:GFP) zebrafish embryos after MO injection. Number of fish for each condition is indicated. (E) Representative examples of sox7/18 double-morphant tg(notch1b-15:GFP) zebrafish at 24 hpf as compared with uninjected controls. Double morphants (dMO) demonstrated reduced EGFP expression in the dorsal aorta and intersomitic vessels. DA, dorsal aorta; PCV, posterior cardinal vein; aISV, arterial intersomitic vessel.
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