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<italic>fbln5</italic> is expressed in the cardiac outflow tract endothelium and its global overexpression partially rescues OFT defects in <italic>alk5</italic> mutants.(A, A’) Whole-mount in situ hybridization for fbln5 expression in the OFT of 36 hpf wild-type larvae (n = 6), combined with immunostaining for kdrl:EGFP (A’). Dotted lines outline the OFT, arrows point to regions with higher staining. (B, B’) Higher magnification of Figure 6C. Fluorescent in situ hybridization for fbln5 expression (white) in the OFT of 36 hpf wild-type embryos. Green, ECs; arrows point to punctae of fbln5 mRNA staining in ECs. (C) Relative mRNA levels of alk5b and fbln5 in 72 hpf alk5+/+, alk5-/-, and Tg(fli1a:alk5b-mScarlet) alk5-/- hearts. Average Ct values are listed in Supplementary file 2. (D) Percentage of OFT expansion in 78 hpf alk5+/+, alk5-/-, and alk5-/- larvae injected with fbln5 mRNA. (E) Percentage of EdU+ ECs in 36 hpf Tg(kdrl:EGFP) alk5+/+ and alk5-/- embryos following injection of fbln5 mRNA. (F) Percentage of SMCs surrounded by Eln2 immunostaining (per sagittal plane) in 75 hpf alk5+/+, alk5-/-, Tg(fli1:fbln5) alk5-/-, and alk5-/- larvae injected with fbln5 mRNA. (C–F) Plot values represent means ± SD; p values from Kruskal-Wallis (C, F) or one-way ANOVA (D, E) tests, compared with the first column (alk5+/+) unless otherwise stated (D). Scale bars: (A, A’) 20 μm; (B, B’) 5 μm.
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