“…We fixed and stained the central nervous system of one adult female Drosophila melanogaster (aged 1-2 days post-eclosion, genotype y,w/w[1118]; +; P{VT025718-Gal4}attP2/P{pBI-UASC-3×MYC-sbAPEX2-dlg-S97}18). Following fixation (2% paraformaldehyde/2.5% glutaraldehyde) and dissection (Tobin et al, 2017), the specimen was reacted with diaminobenzadine (DAB) and H2O2 as described previously (Zhang et al, 2019), but an EM-dense label was not observed in this sample. The dissected central nervous system was then post-fixed and stained with 1% osmium tetroxide/1.5% potassium ferrocyanide, followed by 1% thiocarbohydrazide, a subsequent incubation in 2% osmium tetroxide, then 1% uranyl acetate, followed by lead aspartate (Walton, 1979), then dehydrated with a graded ethanol series.…”