Figure 4

Silencing of cfdp1 expression with a second splice-blocking morpholino. (A) Schematic representation of splicing outcome after microinjection of splice morpholino targeting the exon 2–intron 2 splice junction. The sequenced product shows a frameshift that results in a premature termination codon. (B) Stereoscopic images of representative 72 hpf cfdp1-splice-MO-injected and uninjected control embryos. Black arrows point to the head and heart edema and quantification of phenotypic scoring via GraphPad Prism 9. Data are presented as mean ± SD. n = 50 in each of three independent experiments. Scale bar 150 μm. (C) Max projection of fluorescent images showing that cfdp1 silencing does not affect Notch activity at 72 hpf splice-MO-injected embryos (N = 3, n = 20), while (D) Wnt/β-catenin reporter activity is reduced compared to wild-type control siblings (N = 2, n = 25). AV: Atrioventricular valve; B: bulbus arteriosus; scale bar: 50 μm.