Disruption of calcineurin/NFATc signaling and blocking PRKD activity exhibits a similar phenotype to s411 mutant embryos. Incubation of wild type (A) with 10 μg ml−1 cyclosporine A (CsA) results to a s411 phenotype. s411 heterozygous (B) embryos treated with cyclosporine A, at 2 μg ml−1, a sublethal dose of CsA show phenotype while there is no effect in wild-type embryos (C) when treated at this dose. (D) Treatment of wild-type embryos with a protein kinase D family inAhibitor, 2,3,4,5 -Tetrahydro-7-hydroxy- 1H-benzofuro[2,3-c]azepin-1-one between 30–72 hpf phenocopies s411 mutants showing that this is the critical window of prkd2 activity in the heart. n=20 in each of three independent experiments. Scale bars: 100 μm.
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