Fig. 2

bach2a is essential for developmental angiogenesis in zebrafish embryos.

(A) Confocal images of the primordial hindbrain channel (PHBC, white arrow) of 30-hpf Tg(fli1:EGFP)y1 embryos injected with control MO (10 ng), bach2a MO (3.75 ng), bach2b MO (3.75 ng), bach2b gRNA (125 ng), or vegfcMO (10 ng). Asterisk indicates the absence of PHBC. (B) Percentage of 30-hpf Tg(fli1:EGFP)y1 embryos with intact PHBC formation after injection with control MO (10 ng, nControl MO = 68), bach2a MO (3.75 ng, nbach2a MO = 107; *P< 0.0001), bach2b MO (3.75 ng, nbach2b MO = 48), bach2b gRNA (125 ng, nbach2bgRNA = 42), or vegfc MO (10 ng, nvegfc MO = 35; *P < 0.0001). Error bars, mean ± SEM. (C) Percentage of 30-hpf Tg(fli1:EGFP)y1 embryos with intact PHBC formation after injection with control MO (10 ng, nControl MO = 24) or an increased concentration of bach2b MO (5 ng, nbach2b MO 5ng = 24) or (10 ng, nbach2b MO 10ng = 24). Error bars, mean ± SEM; P > 0.99999. (D) Confocal projection at 30 hpf Tg(fli1:EGFP)y1 of homozygous bach2a mutants (bach2amut−/−) from F2 bach2amut+/− incross. White arrow points at an intact PHBC detected in embryos injected with control MO (10 ng, bach2amut−/− + Control MO) and bach2a MO (3.75 ng, bach2amut−/− + bach2a MO). Asterisk indicates defects in PHBC development after injection with bach2b MO (3.75 ng, bach2amut−/− + bach2b MO) or bach2b gRNA (125 ng, bach2amut−/− + bach2b gRNA). Scale bar, 100 μm. (E) Percentage of randomly selected bach2a+/− F2 incross progeny at 30 hpf with an intact PHBC formation injected with control MO (10 ng, nbach2amut + Control MO = 50; P > 0.99999), bach2a MO (3.75 ng, nbach2amut + bach2a MO = 75; *P < 0.0002), bach2b MO (3.75 ng, nbach2amut + bach2a MO = 55; *P < 0.012) or bach2b gRNA (125 ng, nbach2amut + bach2b gRNA = 137; *P < 0.001). After genotyping, offspring followed the expected Mendelian ratios of inheritance. Error bars, mean ± SEM. Kruskal–Wallis test in panels (B, C, E).