(A) Schematic diagrams of truncated forms of hFz5, RFz2, Plcδ4a and xDvl2. Numbers above the diagram indicate the corresponding amino acid positions. hFz5/RFz2 has N terminal CRD (Cysteine-rich Wnt-binding domain) and Frizzled domain (membrane spanning region of frizzled and smoothened factors). Plcδ4a is comprised of Pleckstrin homology (PH) domain, EF-hand (EF-h) domain, EF-hand (EF-h) like domain, Catalytic TIM barrel domain (incorporating regions of high sequence similarity X and Y) and C2 domain. xDvl2 (GeneID: 108703580) possesses DIX domain (named after Dishevelled and Axin), PDZ domain and DEP domain (named after Dishevelled, Egl-10 and Plekstrin). The truncated forms are indicated by respective amino acid positions. (B) Schematic diagrams of Sec14l3 full length form (FL) and truncated mutants. Numbers above the diagram indicate the corresponding amino acid positions. Sec14l3 includes N terminal CARL-TRIO domain (named after cellular retinaldehyde-binding protein (CRALBP) and TRIO guanine exchange factor), medial Sec14 domain (domain in homologues of a S. cerevisiae phosphatidylinositol transfer protein (Sec14p)) and C terminal GOLD domain (short for Golgi Dynamics domain). ΔN, N terminal CARL-TRIO domain deletion;ΔS, Sec14 domain deletion; ΔGα, Gα domain deletion; ΔG, GOLD domain deletion; ΔNG, N-terminal CARL-TRIO domain and C-terminal GOLD domain deletion. (C) Sec14l3 interacts with Rfz2-CT via its CARL-TRIO and GOLD domains. Domains of Sec14l3 required for interaction with Rfz2-CT in HEK293T cells were tested by co-immunoprecipitation using deletion constructs. TCL, total cell lysate; IP: Immuno-precipitation; WB: western blot. (D) Flag-Sec14l3 interacts with xDvl2-Myc in zebrafish embryos. Embryos were injected with a mixture of 300 pg Flag-Sec14l3 and 200 pg xDvl2-Myc mRNA at 1 cell stage and lysed at 75% epiboly stage for Co-IP assays. (E) Sec14l3 interacts with mDvl2 via its Sec14 domain. (F) xDvl2 interacts with Sec14l3 via its C-terminal region. (G) Flag-Sec14l3 interacts with Myc-Plcδ4a-mCherry in zebrafish embryos. A mixture of 300 pg Flag-Sec14l3 and 200 pg Myc-Plcδ4a-mCherry mRNA was injected into embryos for Co-IP assays. (H) The Sec14l3 CARL-TRIO and GOLD domains interact with Plcδ4a. (I) The Plcδ4a interacts with Sec14l3 via its N-terminal domain.