Figure S1. Sequence alignment of vertebrate h-CaD proteins. Note high degree of conservation in the actin and myosin binding domains (blue lines and green lines, respectively). H_sapiens – Human; G_gallus – Chicken; cald1b – zebrafish paralog (chromosome 25); cald1a – zebrafish paralog (chromosome 4).

Figure S2. Conserved gene synteny surrounding the human CALD1 and zebrafish cald1a and cald1b loci. Schematic depicts the genes surrounding the CALD1 locus on human chromosome 7. Genes with zebrafish orthologs in a comparable location surrounding the cald1a or cald1b loci are depicted in green and orange, respectively. Gene paralogs surrounding both the cald1a and cald1b loci are listed below.

Figure S3. Schematic of a carboxy-terminal splice variant of cald1a. Based on sequencing analysis, the most common cald1a isoform was comprised of 15 exons (top) while a minority splice variant isoform contained an additional exon containing a premature stop codon (bottom).

Figure S4. Average number of intestinal contractions in Cald1a-i4e5 MO and Tg(sm22a:CaDDK51-GFP) larvae in wild type and colourless backgrounds. Contractions were counted using live video imaging, t = 2.5 min. Number of larvae assayed and age (dpf) is indicated for each group. Error bars indicate standard deviation. TgTg(sm22a:CaDDK51-GFP); wt – wild type.

Table S1. Primer sequences for RT-PCR expression analysis. acta2 - smooth muscle actin; myh11a - smooth muscle myosin; fabp2 - fatty acid binding protein 2; vil1l - villin 1 like; fabp6 - fatty acid binding protein 6.

Data S1. Materials and Methods.

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