Fig. 4

Aortic abnormalities in Secisbp2 knockdown zebrafish

A, G and J Representative aortic images, visualized by green fluorescent protein expression as described in Fig. 3 legend, of ventral aorta (VA) and bulbus arteriosus (BA) of zebrafish embryos (5dpf), following zygote injection with control (C), or Secisbp2 morpholinos (MO) or co-injection of Secisbp2 morpholino with Secisbp2 mRNA (Rescue; R). Zygotes were treated (3–120 hpf) with 0.5 mM H₂O₂ or H₂O₂ + 1 mM a-tocopherol (aToc) or H₂O₂ + 1 mM Decyltriphenylphosphonium (TPP) or H₂O₂ + 1 mM MitoQ (G) or treated (from 6 to 120 hpf) with vehicle (DMSO), 100 mM desferrioxiamine (DFO), 10 mM Erastin or 10 mM Erastin + 100 mM DFO (J). The dotted lines A show the positions of VA and BA measurements quantified in (B and C). Each image was repeated independently 15 times with similar results. Scale bar: 50 μm. B, C and F H₂O₂ production (B), membrane lipid peroxidation (C) and p53 and casp3 mRNA expression (F) in VA of 5 pools of 50 embryos at 4 dpf following zygote injection with control (C: grey), or Secisbp2 morpholinos (MO: blue) or co-injection of Secisbp2 morpholino with Secisbp2 mRNA (Rescue; R: yellow). Statistics: One-way ANOVA with adjusted p values (Tukey’s multiple comparison test), each bar represents the mean of n = 5 independent experiments, and error bars represent SEM. D, E, H, I, K and L. VA diameter, measured at positions (1–3) marked in A (D, H, K) or BA diameter (E, I, L) quantified in different groups (5 dpf, n = 15 independent experiments) following zygote injection with control (C), or Secisbp2 morpholinos (MO) or co-injection of Secisbp2 morpholino with Secisbp2 mRNA (Rescue; R) and treated as indicated (0.5 mM H₂O₂; 1 mM α-tocopherol (αToc); 1 mM Decyltriphenylphosphonium (TPP); 1 mM MitoQ). Statistics: Ordinary one-way ANOVA with adjusted p values (Tukey’s multiple comparison test), each bar represents the mean, error bars represent SEM, ns = not significant. Source data are provided as a Source Data file.