Fig. 2: Neuronal DVE-1 is required developmentally for removal of juvenile iAChRs.

a Schematic of dve-1(syb7515) [dve-1::AID::mScarlet] crossed with reSi7 [pan-neuronalpr::TIR1::BFP::AID]. b Timelines of auxin treatments for DVE-1 degradation. c Ventral nerve cord (VNC) images of L4 animals showing DVE-1::AID::mScarlet (red), flp-13pr::ACR-12::GFP (green), and TIR1::BFP::AID (blue) either under control conditions (left) or with continuous auxin treatment from hatch (right). White dashed circle indicates DD1 cell body. Blue arrowhead, intestinal cell. Scale bar, 5 µm. d Quantification of panel (c), scatterplot of average DVE-1::AID::mScarlet fluorescence in DD1 neurons (left) or intestinal cells (right). Each point represents a single animal. Bars indicate mean ± SEM. ****p < 0.0001, ns: not significant, two-tailed students t-test. e Quantification of DD neuron iAChR clusters in the L4 stage dorsal nerve cord for control and dve-1::AID::mScarlet animals under treatment conditions described in panel (b). Each point represents a single animal. Bars indicate mean ± SEM.,****p < 0.0001, ***p < 0.001, two-way ANOVA with Tukey’s multiple comparison. f Confocal images of ACR-12::GFP clusters in the dorsal nerve cord of L4 stage dve-1::AID::mScarlet animals either under control conditions or with continuous auxin treatment. Scale bar, 5 µm. g Top, scatterplot of LEV-10::GFP dorsal/ventral fluorescence intensity ratio measurements per corresponding 25 µm regions of dorsal and ventral nerve cord expressed as dorsal/ventral fluorescence ratio −1. Bars indicate mean ± SEM. ***p < 0.001, ns: not significant two-tailed student’s t-test. Bottom, confocal images of LEV-10::GFP from the dorsal nerve cord before (pre-remodeling) and after (post-remodeling) the L1/L2 transition. Scale bar, 5 µm. NATF DD LEV-10 indicates tissue-specific labeling of endogenous LEV-10 by split-GFP³⁶. Each point represents a single animal. h Top, confocal images of the dorsal and ventral process from L4 stage wild-type and dve-1(uf171) mutants co-expressing the mCherry::RAB-3 synaptic vesicle (SV) and ACR-12::GFP (iAChR) markers in DD neurons. Scale bar, 5 µm. Bottom, quantification of iAChR clusters (left) and SV puncta (right) in dorsal and ventral processes of L4 stage DD neurons for wild-type and dve-1(uf171) mutants. Each point represents a single animal. Bars indicate mean ± SEM. ****p < 0.0001, ns: not significant, two-way ANOVA with Tukey’s multiple comparison test.