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Fig. 7
Putative N-glycosylation sites of Fz8 receptor are important for Wnt8 signaling. (A) Schematic representation of putative N-glycosylation sites in zebrafish Fz8a receptor. Abbreviations. CRD, Cystein rich domain; ICD, Intercellular domain; SP, Signal peptide; TM, Transmembrane domain. (B–F) HEK293 cells were transfected with indicated Fz8 construct and overexpressed Fz8 protein is analyzed by anti-Myc immunoblotting (B) or anti-Myc immunostaining (green) (C–F). For panels C–J, nuclei are counter-stained with Hoechst 33342 (blue). Full-length blots are presented in Supplementary Fig. S19. (G—J) Immunostaining of Myc (green) in embryos (6 hpf) injected with mRNA for indicated wild type or mutant fz8a-myc. (K) Embryos were injected with indicated combinations of wnt8 and fz8 mRNAs. Left panel: Resulting phenotypes are classified from Class I (normal) to Class 4 (most severe). Right panel: Phenotype frequencies of embryos injected with indicated combinations of mRNAs.