Figure 7.
unc-73 pathway negatively regulates klp-7 to promote the growth of the anterior process. (A) Phosphorylation sites of PAK on KLP-7. (B) The processed images of a worm expressing Ppak-1-PAK-1::GFP (shrEx439) and Pmec-4-mCherry (tbIs222) for seeing PLM-specific localization of PAK-1. (C) The relative intensity of PAK-1::GFP with respect to mCherry from the ROIs shown in B. Ant, anterior; Pst, posterior. (D) Image of the PLA between α-GFP and α-KLP-7 antibodies in a worm expressing PAK-1::GFP. Arrowheads indicate the PLA foci in the PLM anterior process. (E) The count of PLA foci per micrometer of PLM processes shown in D. (F) Confocal images of a PLM neuron in unc-73(0) and unc-73(0);klp-7(0). (G and H) Normalized anterior (G) and posterior (H) lengths of the PLM in the single mutant of unc-73 pathway or combination with either klp-7(0) or a transgene shrEx266 (Punc-86-KLP-7S546E S555E) expressing a B-type inactive version of KLP-7. For C, E, G, and H, N = 3–4 independent replicates; n (number of neurons) = 42–68. (I) Confocal images of a PLM neuron in lin-17(0) coexpressing the A-type hyperactive Punc-86-KLP-7T549E and B-type inactive Punc-86-KLP-7S546E S555E versions of KLP-7 using shrEx197 or expressing the activated version of CED-10, CED-10G12V (shrEx294). (J and K) Normalized anterior (J) and posterior (K) lengths of the PLM in lin-17(0) background expressing various mutant forms of KLP-7 and CED-10. N = 3–4 independent replicates; n (number of neurons) = 32–115. (L) Pathway showing how UNC-73/CED-10 regulates KLP-7. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ANOVA with Tukey’s multiple comparison test. Error bars represent SEM.

unc-73 pathway negatively regulates klp-7 to promote the growth of the anterior process. (A) Phosphorylation sites of PAK on KLP-7. (B) The processed images of a worm expressing Ppak-1-PAK-1::GFP (shrEx439) and Pmec-4-mCherry (tbIs222) for seeing PLM-specific localization of PAK-1. (C) The relative intensity of PAK-1::GFP with respect to mCherry from the ROIs shown in B. Ant, anterior; Pst, posterior. (D) Image of the PLA between α-GFP and α-KLP-7 antibodies in a worm expressing PAK-1::GFP. Arrowheads indicate the PLA foci in the PLM anterior process. (E) The count of PLA foci per micrometer of PLM processes shown in D. (F) Confocal images of a PLM neuron in unc-73(0) and unc-73(0);klp-7(0). (G and H) Normalized anterior (G) and posterior (H) lengths of the PLM in the single mutant of unc-73 pathway or combination with either klp-7(0) or a transgene shrEx266 (Punc-86-KLP-7S546E S555E) expressing a B-type inactive version of KLP-7. For C, E, G, and H, N = 3–4 independent replicates; n (number of neurons) = 42–68. (I) Confocal images of a PLM neuron in lin-17(0) coexpressing the A-type hyperactive Punc-86-KLP-7T549E and B-type inactive Punc-86-KLP-7S546E S555E versions of KLP-7 using shrEx197 or expressing the activated version of CED-10, CED-10G12V (shrEx294). (J and K) Normalized anterior (J) and posterior (K) lengths of the PLM in lin-17(0) background expressing various mutant forms of KLP-7 and CED-10. N = 3–4 independent replicates; n (number of neurons) = 32–115. (L) Pathway showing how UNC-73/CED-10 regulates KLP-7. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ANOVA with Tukey’s multiple comparison test. Error bars represent SEM.

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