Figure 1. Development of Q neuroblasts in C. elegans
Schematic diagrams of Q neuroblast migration and the Q lineages. (A) QR and QL neuroblasts are born at similar positions at the right and left side of the worm body and their descendants migrate to the anterior and posterior, respectively, during development.
(B) The QL and QR display identical patterns of cell division.
Figure 2. Canonical Wnt signaling
Schematic model of -catenin-dependent Wnt signaling. C. elegans homologs required for QL.d migration are noted on the right.
Figure 3. vang-1 antagonizes Wnt pathwaysignaling at the level of the Frizzled receptor
(A) Schematic diagram of the PVM migration process. The PVM positions are defined as the ratio of the anterior-to-PVM length (a) to the anterior-posterior body length of the animal (b). (B) PVM positions in the wild type and the Wnt pathway mutants. PVM positions are labeled by the transgene zdIs5[Pmec-4::gfp]. n.s., not significant. ***, p <
0.005, unpaired t test. Numbers of neurons scored are indicated behind the genotypes.
Error bars represent the S.E.M.
Figure 4. Loss of vang-1 function recues PVM migration defect in mig-1 mutant PVM positions in the wild type and indicated mutants. PVM positions are labeled by the transgene zdIs5[Pmec-4::gfp]. *, p < 0.05, ***, p < 0.005, unpaired t test. Numbers of neurons scored are indicated behind the genotypes. Error bars represent the S.E.M.
Figure 5. The vang-1 mutation restores mab-5 expression in the mig-1 mutant Epifluorescence images of QL.a and QL.p in animals expressing Pmab-5::GFP. The Q.d cells are labeled by myr::mCherry in indicated genotypes. Scale bar = 10µm.
Figure 6. Quantification of mab-5 expression
Quantification of mab-5 expression in indicated genotypes. n.s., not significant. ***, p <
0.005, Fisher’s exact test. Numbers are neurons quantified for Pmab-5::GFP intensity.
Figure 7. vang-1 functions autonomously to antagonize Wnt signaling in QL.d migration
Quantification of PVM positions in indicated genotypes. ##, p < 0.01; ###, p < 0.005, F test. **, p < 0.01, unpaired t test. Numbers of neurons scored are indicated behind the genotypes. Error bars represent the S.E.M.
Figure 8. vang-1 antagonizes Wnt signaling in QR.d migration
(A) Diagram of the AVM migration process. The AVM positions are defined as the ratio of the anterior-to-AVM length (a) to the anterior-posterior body length of the animal (b).
(B, C) Quantification of AVM positions in the wild type and Wnt pathway mutants. The AVM positions are labeled by the transgene zdIs5[Pmec-4::gfp]. n.s., not significant. ***, p < 0.005, unpaired t test. Numbers of neurons scored are indicated behind the genotypes.
Error bars represent the S.E.M.
Figure 9. vang-1 antagonizes Wnt signaling in HSN migration
(A) Diagram of HSN migration during larval development. The HSN positions are defined as the ratio of the vulva-to-HSN length (a) to the vulva-tail body length of the animal (b). (B) Quantificatiion of HSN positions in the wild type and Wnt pathway mutants. The HSN positions are labeled by the transgene mgIs42[Ptph-1::gfp]. ***, p <
0.005, unpaired t test. Numbers of neurons scored are indicated behind the genotypes.
Error bars represent the S.E.M.
Figure 10. vang-1 antagonizes Wnt signaling autonomously in HSN migration (A, B) Quantificatiion of HSN positions in the wild type and Wnt pathway mutants. The HSN positions are labeled by the transgene mgIs42[Ptph-1::gfp] in the wild type, mig-1 and mig-1;vang-1, and by twnEx199[Punc-86::gfp::vang-1] in the mig-1 mom-5 and mig-1 mom-5; van.g-1. n.s., not significant. ***, p < 0.005, unpaired t test. Numbers of neurons scored are indicated behind the genotypes. Error bars represent the S.E.M
Figure 11. vang-1 antagonizes Wnt signaling in ALM polarity
(A) Diagram of the ALM polarity phenotypes. (B) Quantification of ALM polarity defects in indicated genotypes. The ALM neurons are marked by the transgene zdIs5[Pmec-4::GFP]. n.s., not significance. *, p < 0.05; two proportion z test. Numbers are neurons scored.
Figure 12. Overexpression of vang-1 causes polarity defects in the ALM
Quantification of ALM polarity defects in indicated genotypes. **, p < 0.01, ***, p <
0.005, two proportion z test. Numbers are neurons scored.
Figure 13. arr-1 acts downstream of vang-1 to antagonize Wnt signaling in PVM migration
(A, B) Quantification of PVM positions in indicated genotypes. n.s., not significant. ***, p < 0.005; unpaired t test; ###, p < 0.005, F test. Numbers of neurons scored are indicated behind the genotypes. Error bars represent the S.E.M.
Figure 14. arr-1 acts downstream of vang-1 to antagonize Wnt signaling in AVM migration
(A, B) Quantification of PVM positions in indicated genotypes. *, p < 0.05; ***, p <
0.005, unpaired t test. Numbers of neurons scored are indicated behind the genotypes.
Error bars represent the S.E.M.
Figure 15. arr-1 acts in the same pathway with vang-1 to antagonize Wnt signaling in HSN migration
Quantification of HSN positions, which are labeled by the transgene mgIs42[Ptph-1::gfp].
n.s., not significant. ***, p < 0.005, unpaired t test. Numbers are neurons scored. Error bars represent the S.E.M.
Figure 16. arr-1 acts downstream of vang-1 to antagonize Wnt signaling in ALM polarity
Quantification of the ALM polarity defects in indicated genotypes. *, p < 0.05, two proportion z test. Numbers are neurons scored.
Figure 17. vang-1 promotes MOM-5 endocytosis.
(A) Representative confocal images of animals expressing MOM-5::GFP fed with control or vang-1 RNAi bacteria. The MOM-5 transgene is zuIs145. The Q.d cells are labeled by rdvIs1. (B) Quantification of membrane MOM-5::GFP signal as a ratio to MOM-5::GFP signal in the cytosol. Error bars represent the S.E.M.
Figure 18. Frizzleds and VANG-1 protein structures
Schematic diagrams of Frizzleds and VANG-1 protein structures. FL, full-length; TM, transmembrane domain.
Figure 19. MOM-5 forms a complex with VANG-1
Co-immunoprecipitation of MOM-5::FLAG and HA::VANG-1 expressed in cultured HEK293 cells.
Figure 20. MIG-1 formss complexes with VANG-1 (1-281)
Co-immunoprecipitation of MIG-1::FLAG and HA::VANG-1 (1-281) expressed in the HEK293 cells.
Figure 21. MOM-5 forms complexes with VANG-1 (1-281)
Co-immunoprecipitation of MOM-5::FLAG and HA::VANG-1 (1-281) expressed in the HEK293 cells..
Figure 22. MIG-1 forms a complex with VANG-1 and DSH-2
Co-immunoprecipitation of MIG-1::FLAG, HA::VANG-1 and DSH-2::MYC expressed in the HEK293 cells.
Figure 23. DSH-2 is not part of the MOM-5-VANG-1 complex
Co-immunoprecipitation of MOM-5::FLAG, HA::VANG-1 and DSH-2::MYC expressed in the HEK293 cells.
Figure 24. Model of VANG-1 modulation of Wnt signaling.