UNC-57/Endophilin is enriched in males at their cholinergic motor neuron terminals. (A) Schematic illustration of split GFP complementation experimental design. Seven copies of the split GFP11 were inserted into the C-terminal of unc-57 (T04D1.3a) genomic loci by CRISPR-Cas9 system. The split GFP1-10 was expressed in the B-type cholinergic and GABAergic motor neurons by unc-129 and unc-25 promoters, respectively. (B–D) The puncta fluorescence intensities and densities, marked by the cholinergic synaptic UNC-57::split GFP in dorsal nerve cord axons in hermaphrodites (Herm.) and males (Male). Representative images (B), mean puncta intensity (C), and density (D) are shown. (E–G) The puncta fluorescence intensities and densities, marked by the GABAergic synaptic UNC-57::split GFP in dorsal nerve cord axons in hermaphrodites (Herm.) and males (Male). Representative images (E), mean puncta intensity (F), and density (G) are shown. In B and E, Scale bars, 10 μm. In C, D, F, and G, data are presented as mean values ± SEM. Ns represent the number of animals tested. Two-tailed and unpaired Student’s t test. **P < 0.01, n.s. not significant.