Fig. 6: The deSUMOylating peptidase ULP-4 regulates DVE-1 nuclear localization. | Nature Communications

Fig. 6: The deSUMOylating peptidase ULP-4 regulates DVE-1 nuclear localization.

From: The homeodomain transcriptional regulator DVE-1 directs a program for synapse elimination during circuit remodeling

Fig. 6: The deSUMOylating peptidase ULP-4 regulates DVE-1 nuclear localization.The alternative text for this image may have been generated using AI.

a Fluorescence intensity of nuclear DVE-1::GFP in DD motor neurons at 10, 18, 22 and 50 h after hatch. Nuclear DVE-1::GFP is organized in discrete foci and increases through development. 10 h n = 10, 18 h n = 7, 22 h n = 8, 50 h n = 9. Each time point indicates mean ± SEM from at least three independent experiments. Inset, representative image of nuclear DVE-1::GFP in DD motor neuron 18 h after hatch (nucleus labeled by NLS::mCherry). White dashed line outlines nucleus. Scale bar, 3 µm. b Confocal fluorescence images (left) of DVE-1::GFP in DD GABAergic motor neurons of L1 stage wild-type, ulp-4(lf) mutant, and K327R;ulp-4(lf) double mutants. ulp-4(lf) disrupts DVE-1::GFP nuclear localization. White dashed line outlines nucleus. Scale bar, 3 µm. Right, line scan of nuclear DVE-1::GFP fluorescence intensity in DD motor neurons of wild-type (green) n = 16, ulp-4(lf) (blue) n = 11 and DVE-1(K327R);ulp-4(lf) double mutants (yellow) n = 8. Solid line represents mean, shading represents standard deviation of fluorescence. c Scatterplot (left) of the peak nuclear DVE-1::GFP fluorescence intensity in DD motor neurons. Each point represents a single DD nucleus. Imaged at least 2 DD cells per animal at L1 stage. Wild type: n = 16, ulp-4(lf): n = 11, K327R;ulp-4(lf): n = 8. Bars indicate mean ± SEM. ****p < 0.0001, one-way ANOVA with Tukeys multiple comparisons test. d Fluorescent confocal images of iAChR clusters in the dorsal processes of L4 stage DD neurons for genotypes indicated. Scale bar, 5 µm. e Quantification of iAChR clustering at L4 stage for the genotypes indicated. Bars indicate the percentage of L4 stage animals where dorsal iAChRs have been completely removed. ****p < 0.0001, *p < 0.05, ns: not significant, two-tailed Fischer’s exact test with Bonferroni correction. 1/2 pan-neuronal lines, 2/2 GABA lines, and 0/2 intestinal rescue lines restored proper removal of dorsal iAChRs by L4. f Scatterplot of average iAChR fluorescence intensity per 25 µm in the dorsal nerve cord at L4 stage for the genotypes indicated. Each point represents a single animal. Bars indicate mean ± SEM. ****p < 0.0001, ***p < 0.001, **p < 0.01, one-way ANOVA with Tukey’s multiple comparisons test.

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