Figure 2. The hpo-30/Claudin, tiam-1/GEF, and act-4/Actin act genetically in the Menorin pathway.

(A – D) Fluorescent images of PVD (left panels, visualized by the wdIs52 [F49H12.4p::GFP] transgene) and schematics (right panels) of the genotypes indicated. The control image is identical to Figure 1A and shown for comparison only. Details on alleles of individual genes and images of other alleles are shown in Figure 1—figure supplement 1. Scale bar indicates 20 µm. (E – G) Quantification of 2°, 3°, and 4° branch numbers per 100 µm anterior to the PVD cell body. Data for additional single and double mutants of the Menorin pathway as well as the average length and aggregate length of secondary, tertiary, and quaternary branches are shown in Figure 2—figure supplement 2. All alleles used are molecular or genetic null alleles (Material and methods for details). Data for control, lect-2, sax-7, mnr-1 and dma-1 mutant animals is identical to data from Díaz-Balzac et al. (2016) and shown for comparison only. For raw data see Figure 1—source data 1. Data are represented as mean ± SEM. Statistical comparisons were performed using one-sided ANOVA with the Tukey correction. Statistical significance is indicated (ns, not significant; *, p<0.05; **, p<0.01; ***, p<0.001; ****, p<0.0001). n = 20 animals for all genotypes.

Figure 2—figure supplement 1. Effects of hpo-30/Claudin, tiam-1/GEF and act-4/Actin on localization of LECT-2:mNG, SAX-7::GFP and DMA-1::GFP.

(A) Fluorescent images of animals expressing GFP in FLP neurons (muIs32 [Pmec-7::GFP]) in the indicated genotypes. Anterior is to the left and scale bars indicate 20 µm. (B – D) Fluorescent images of animals expressing a LECT-2::mNeonGreen fusion at endogenous levels (a functional knock in (lect-2(dz249) [lect-2::mNG^3xFlag] (Díaz-Balzac et al., 2016)) (B), a functional fosmid-based SAX-7::GFP reporter (ddIs290) (C), and a functional DMA-1::GFP reporter (D) (Liu and Shen, 2011). Arrowheads indicate the localization of LECT-2::mNG or SAX-7::GFP in a stripe at edge of the lateral hypodermis, where hypodermis and muscle abut. The control image (B) is identical to Figure 4B and shown for comparison only. A green box indicated enrichment of DMA-1::GFP at the tip of dendrites of act-4(dz222) mutant animals. (E) Quantification of DMA-1::GFP puncta in the genotypes indicated. n = 14 for control, n = 12 for tiam-1(tm1556), and n = 11 for act-4(dz222). For raw data see Figure 1— source data 1. Statistical significance is indicated as Data are represented as mean ±SEM. Statistical comparisons were performed using one-sided ANOVA with the Tukey correction. Statistical significance is indicated (ns: not significant, **p<0.05, ***p<0.005, ****p<0.0005).

Figure 2—figure supplement 2. The Genetics of hpo-30/Claudin, tiam-1/GEF, and act-4/Actin.

(A – H) Fluorescent images of animals expressing GFP in PVD neurons (wdIs52 [F49H12.4p::GFP]) in the indicated genotypes. Anterior is to the left and scale bars indicate 20 µm. (I – K) Quantification of the aggregate length of secondary, tertiary, and quaternary branches 100 µm anterior to the PVD cell body. Data are represented as mean ± SEM. Statistical comparisons were performed using one-sided ANOVA with the Tukey correction. For raw data see Figure 1— source data 1. Statistical significance is indicated as Statistical significance is indicated (ns: not significant, **p<0.01, ***p<0.001, ****p<0.0001). n = 20 for all samples. (L – Q) Quantification of the number (KM,O) or aggregate length (LN,P) of secondary, tertiary, and quaternary branches 100 µm anterior to the PVD cell body. Some data in (KM,O) is identical to data in Figure 2 and shown for comparison only. For raw data see Figure 1— source data 1. Statistical significance is indicated as Data are represented as mean ± SEM. Statistical comparisons were performed using one-sided ANOVA with the Tukey correction. Statistical significance is indicated (ns: not significant, **p<0.01, ***p<0.001, ****p<0.0001). n = 20 for all samples.