(A) MO injections were performed with wild-type embryos to generate single prkcι, prkcζ, and double prkcι/ζ knockdowns, and morphants were scored at 72 hpf for gross morphology compared to embryos injected with control double prkcι/ζ mismatch MOs. (Left) Percentage distributions of three classes of phenotypes, categorized as wild-type, mild, and severe. These categories were based on curled tail, edema, and patchy eye pigmentation (Right, lateral views of live representative embryos). (B) Confocal imaging of IF analysis during tubulogenesis in wild-type, prkcι, prkcζ and prkcι/ζ morphant nephrons at the 36 hpf stage. Single morphants showed expression and proper localization of Prkcι/ζ and p-ERM to the apical surface, whereas Prkcι/ζ and p-ERM were absent in the double prkcι/ζ morphants. Na+/K+ ATPase was basolateral in wild-type embryos and single prkc morphants, but diffuse in the double prkcι/ζ morphants. Scale bars, 5 μm. (C) WISH analysis of podocyte and PCT development in wild-type control embryos and single prkc morphants using wt1a and slc20a1a, respectively. prkcζ morphants had midline clusters of podocytes, similar to wild-type embryos, while prkcι morphants had scattered clusters of podocytes that failed to migrate to the midline. PCT coiling morphogenesis was normal in prkcζ morphants and wild-type control embryos, while prkcι morphants had disrupted PCT coiling morphogenesis. (Left columns, lateral view; right columns, dorsal view).