Fig. 7: MADD-4/Punctin controls the synaptic localization of the LIN-2-FRM-3 complex.

a, b Confocal detection of FRM-3A-GFP expressed in the muscle of wild-type animals and in mutants lacking the MADD-4 short isoform: madd-4B(0); the long isoforms: madd-4L(0); or all isoforms: madd-4(0). FRM-3A-GFP fluorescence intensity was normalized to the wild type. Box plot as in Fig. 1e. One-way ANOVA followed by Turkey’s multiple comparison tests. ***p < 0.001. n.s., not significant. c, f Confocal imaging of the dorsal cord of wild-type of animals expressing FRM-3A-GFP in muscle and SNB-1-BFP in GABA motoneurons to label presynaptic GABAergic boutons. The fluorescence profiles indicate FRM-3A-GFP and SNB-1-BFP fluorescence intensities along the nerve cord from the pictures above. f Mander’s overlap correlation was used to calculate the percentage of FRM-3A-GFP signal that overlaps with the SNB-1-BFP signal. Box plot as in Fig. 2d. Kruskal–Wallis test followed by Dunn’s post test. ***p < 0.001. n.s, not significant. g, h GFP-LIN-2A was expressed in the muscle of wild-type and mutant animals and SNB-1-BFP was used to label GABAergic boutons (images are shown in Fig S3). g GFP-LIN-2A fluorescence intensity was normalized to the wild type. Box plot as in Fig. 1g. One-way ANOVA followed by Turkey’s multiple comparison tests. ***p < 0.001. n.s, not significant. h Mander’s overlap correlation indicates the percentage of GFP-LIN-2A signal that overlaps with the SNB-1-BFP signal. Box plot as in Fig. 2d. Kruskal–Wallis test followed by Dunn’s post test. **p < 0.01. n.s, not significant. Scale bars = 10 μm.