Generation of cav1-KO by CRISPR/Cas9 editing and transcriptional analysis. (a) Schematic representations of the two cav1 transcripts—cav1a and cav1b. Target site of the guide RNA is indicated by a red arrowhead. ex, exon. (b) Identified genetic mutations. Red dashed lines indicate deletions, red uppercase letters insertions, and black lowercase letters silent mutations. bp, base pairs. (c) Cav1 domain organisation and the predicted effect of the mutations on the protein. The novel amino acids are in red, x indicates a stop codon. (d) qPCR analysis of caveolae-related genes in two-day post fertilisation embryos. Mean ± s.d., t-test, **P < 0.01. (e–j) Immunostaining of Cav1a in 7 dpci hearts. (f, g, i, j) Higher magnification of the dashed boxes in (e) and (h), respectively. (k–p) Cav1 (Cav1 and Cav1b) immunolabelling of 7 dpci hearts. (l, m, o, p) Higher magnification of the dashed boxes in (k) and (n), respectively. Dotted areas in (h) and (n) mark the valves in the mutants. Scale bars: 100 μm in (e), (h), (k), (n); 50 μm in other panels.
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