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Fig. S2
alk1 MO efficacy and effects of DAPT on MC emergence. (A) The schematic model of alk1 MO target site (left). alk1 MO targets the border between exon 6 and intron 7, resulted in the skipping of exon 6 (*) or the failure of removal of intron 7 (**) as shown in the right PCR result. (B) Confocal stack images of the central arteries of 60 hpf TgBAC(pdgfrb:GFP);Tg(fli1a:Myr-mCherry) embryos injected with 3 ng alk1 MO. Boxed regions are enlarged to the bottom. Note that pdgfrbhigh MCs were not detected around central arteries but fli1a:Myr-mCherry-positive ECs became positive for pdgfrb:GFP (arrows). (C) Representative confocal stack images of the trunk vasculature of 72 hpf TgBAC(pdgfrb:GFP);Tg(fli1a:Myr-mCherry) larva treated with 100 μM DAPT from 24-36 Development: doi:10.1242/dev.165589: Supplementary information Development • Supplementary information hpf. Boxed regions are enlarged to the right. The quantitative analysis of DA-MC coverage is shown below the images. Note that DAPT treatment before the DA-MC emergence period (36 hpf-) did not affect the DA-MC emergence. (D) Representative confocal stack images of the trunk vasculature of 72 hpf TgBAC(pdgfrb:GFP);Tg(fli1a:Myr-mCherry) larva treated with 5 μM LY411575 from 24-72 hpf. (E) Confocal stack images of ISVs in the 96 hpf TgBAC(pdgfrb:GFP);Tg(fli1a:Myr-mCherry) embryos treated with DMSO (control), 100 µM DAPT from 48 hpf to 96 hpf, or DMSO from 48 hpf to 72 hpf and subsequent DAPT from 72 hpf to 96 hpf, as shown in the schematic model on the right top. Arrowheads indicate MCs in ISVs. a, aISV. v, vISV, The percentage of aISV covered by pdgfrbhigh MCs and the number of MCs in one aISV with MCs, as observed in the right images, are shown on the right. In embryos treated with DAPT from 48 hpf, only two out of 18 embryos showed the aISV with pdgfrbhigh MC.