Zebrafish Protocols for Vascular Development

Zebrafish were maintained according to standard protocols (59 ). Zebrafish were kept at 28.5 ^∘C in a 14-h light and 10-h dark cycle. After natural spawning, embryos were collected and raised at 28.5 ^∘C in 0.5 × E2 medium containing methylene blue (egg water). To avoid pigmentation, embryos were changed to 0.003% N-Phenylthiourea (P7629, Sigma) in egg water at 1 dpf. The following strains were used: Tg(flk1:eGFP) ${}^{s843}$ (31 (link)), Tg(flk1:NLS-Eos) ${}^{\mathit{ncv}6}$ (32 (link)), Tg(Tp1:eGFP) ${}^{um14}$ (60 (link)), Tg(hsp70l:NICD-eGFP) (61 (link)), Tg(fli1:DsRed) ${}^{um13}$ (62 (link)), Tg(flk1:loxP-DsRedx-loxP-eGFP) (63 (link)), mib ${}^{ta52b}$ (64 (link)), Tg(etv2:NLS-d2eGFP) $\mathit{hkz}037$ , Tg(hsp70l:etv2-P2a-mCherry) $\mathit{hkz}038$ , Tg(etv2:mCherry-T2a-CreER ${}^{T2}$
) $\mathit{hkz}039$ , Tg(flk1:loxP-DsRedx-loxP-eGFP) $\mathit{hkz}040$ , and Tg(flk1:runx1-P2a-GFP) $\mathit{hkz}041$ . All animal experiments were conducted according to the guidelines of the Animal and Plant Care Facility and approved by the Animal Ethics Committee of the Hong Kong University of Science and Technology.