A. Heat map comparison of gene expression between control siblings (control) and partially paralyzed sofa potato mutants (sop^−/−). Omc5 gene is marked with an arrowhead. B. ddPCR of omc5 transcript in control embryos (control) and paralyzed mutants (sop^−/−) at 5 dpf. C. Diagram displaying protein products of omcin 1-15 along with human uromodulin, human GP2, and human uromodulin-like. Domains/motifs are indicated with colored squares. D. A map of ∼400 KB region on Chromosome 12 where a cluster of omcin genes are located. E. Phylogenetic tree for the amino acid sequences of the zebrafish omcin genes, zebrafish uromodulin-like gene, human uromodulin gene, human GP2 gene, and human uromodulin-like gene. Numbers in the nodes of the tree indicate bootstrap values.

A. Amino acid sequence of omcin gene products surrounding the epitope used for the generation of antibody (boxed in blue). 6 of omcin genes including omc5 have the identical sequence for the epitope, while additional 3 genes have close sequences. B. Western blot of the Omcin antibody on5dpf embryos. The strongest signal was observed around 180 kDa (arrowhead).C. Antibody signals observed near cloaca in control and sop^−/− embryos (arrows).D. Double staining in wild type embryos of Omcin antibody and cdh17. Signals indicated by arrows overlapped in the distal pronephros.E. Omcin antibody staining in slices of control and sop^−/−embryos at 5 dpf. Signals in the gut (arrows) and the pronephros (arrowheads) are indicated. Scale bars, 50 μm.

A. A time course of E. coli incubation experiment. Embryos were cultured in solution with or without E. coli from 1 dpf through 7 dpf. Surviving embryos were counted each day. B. Survival of wild type embryos were plotted against time in E. coli (−) or E. coli (+) solution. C. ddPCR of omc5 in 5 dpf wild type embryos cultured in E. coli (−) or E. coli (+) solution. D. Survival of sop^−/−embryos plotted against time in E. coli (−) or E. coli (+) solution. E. Neutrophils marked with GFP at 4 dpf embryos in E. coli (−) or E. coli (+) solution. 4 embryos were observed for each group and representative images are shown.

A. Diagram of omc5 gene with exons indicated as blue boxes. Locations of nonsense mutation and the sequence encoding the epitope are indicated with arrows. B. Omcin antibody staining in omc5^−/− embryos. C. Survival of omc5^−/− embryos in germ-free solution (GF) and non germ-free solution (non-GF). D. Survival of wild type embryos in germ-free solution (GF) and non germ-free solution (non-GF). E. Survival of wild type and omc5^−/− embryos in E.coli (+) solution. F. myd88 transcript measured by ddPCR in wild type (control) and omc5^−/− embryos after incubation in control solution (PAM(−)), Peptidoglycan (PG), Lipopolysaccharide (LPS) and Flagellin (Fla). G. 10 adult fish were housed in still water for 2 days. Wild type fish remained unaffected, while all omc5^−/− fish died.

A. A maximum-likelihood phylogenetic tree of the zebrafish omcins and omcin-like genes found in the 6 fishes (goldfish, electric eel, piranha, milkfish, northern pike, channel catfish, European anchovy, and mummichog). Numbers in each tree node indicate bootstrap values (only the values > 50 % are shown). B. Chromosomal locations of the omcins in zebrafish and omcin-like genes in the 8 fishes examined. Phylogenetic relationship of these fishes is also shown. Vertical lines indicate the start codon positions. Transcriptional orientations are shown by colors (red: forward strand, black: reverse strand).