Extended Data Fig. 7: Expression pattern of PU-splitGAL4 lines.

a, Representative images from PU-2 > mCD8-GFP females. Left: ventral nerve cord (one of 13 flies) stained with anti-GFP (membrane of PU neurons, green) and nc82 (synaptic neuropil, magenta). Right: lower reproductive tract (one of eight flies) stained with anti-GFP (green) and phalloidin (muscle f-actin, gray); autofluorescence abdominal cuticle (red). In addition to consistent labeling of the PU neurons (PU-1, 2.1 ± 0.4 ventral afferents per side, n = 17 sides in thirteen flies; PU-2, 2.0 ± 0.6; n = 11 sides in eight flies; mean ± s.d.), both lines exhibited inconsistent labeling in a small number of peripheral neurons that project to the dorsal abdominal neuromere (PU-1, 1.2 ± 0.6 dorsal afferents per side; PU-2, 0.5 ± 0.6; mean ± s.d.). Here and in b-d, scale bar, 50 μm. b, Representative images of the brain from PU-1>mCD8-GFP (left, one of three flies) and PU-2>mCD8-GFP (right, one of two flies) females, stained with anti-GFP (green) and nc82 (magenta). c, Representative images from hs-FLP; PU-2>(FRT.stop)CsChrimson-mVenus females (one of two flies) where stochastic labeling resulted in only a single PU neuron being labelled (Gordon and Scott, Neuron 61, 373–384 (2009)). Top two images and right image: ventral nerve cord stained with anti-GFP (mVenus-expressing PU neurons, green) and nc82 (magenta). Bottom two images: lower reproductive tract stained with anti-GFP (green) and phalloidin (muscle f-actin, gray); autofluorescence, abdominal cuticle (red); white triangle, PU cell body; u, uterus. Right: lateral projection of the ventral nerve cord after registration with a template; V, ventral; P, posterior. d, Representative images of the abdominal neuromere (top row, one of three flies) and lower reproductive tract (middle, bottom rows, one of three flies) from PU-2>DenMark, synaptotagmin-GFP females, stained with anti-DsRed (dendrites, red) and anti-GFP (synaptic vesicles, green)^52,53.