FIGURE 2.

FLP³³⁵ restricted labeling of fru-Gal4. (A) Expression profile of FLP³³⁵ fru-GAL4 UAS>stop>mCD8::GFP showing sexually dimorphic projections in the glomeruli from fru ORNs (DA1, VA1v, VL2a), projections in the antennal mechanosensory motor complex (AMMC) from fru JONs, sexually dimorphic projections in the prothoracic neuromere from fru LAN1 in the forelegs, and sexually dimorphic arbors in the abdominal neuromere (Abg). Tissues were stained with anti-mCD8 (green) and anti-nC82 (magenta). Scale Bar = 50 μm. (B) Expression profile of fru-GAL4, UAS>stop>mCD8::GFP, FLP³³⁵, tsh-GAL80. The addition of tsh-Gal80 transgene resulted in less expression in the ventral cord, particularly at the abdominal ganglion. (F,G) Magnified image of abdominal ganglion showing fru neurons. Immunofluorescence is reduced with the introduction of tsh-Gal80 (G). Tissues were stained with anti-mCD8 (green) and anti-nC82 (magenta). Scale Bar = 50 μm. (C–E) Effect of silencing FLP³³⁵ restricted labeling of fru-Gal4 with and without tsh-GAL80. (C) Percentage of successful matings in 1 h (FLP³³⁵, fru>TNTin: n = 33; FLP³³⁵, fru>TNT: n = 48; FLP³³⁵, fru>TNTin, tsh-GAL80: n = 24; FLP³³⁵, fru>TNT, tsh-GAL80: n = 40). ^∗∗∗p = 0.0001, ^∗∗∗p = 0.0004 by Fisher’s exact test. (D) Copulation latency (central line indicates the median; FLP³³⁵, fru>TNTin: n = 27; FLP³³⁵, fru>TNT: n = 10; FLP³³⁵, fru>TNTin, tsh-GAL80: n = 22; FLP³³⁵, fru>TNT, tsh-GAL80: n = 28). ^∗∗∗p = 0.0002, n.s. = not significant by Mann–Whitney test. (E) Copulation duration (central line indicates the median; FLP³³⁵, fru>TNTin: n = 27; FLP³³⁵, fru>TNT: n = 16; FLP³³⁵, fru>TNTin, tsh-GAL80: n = 22; FLP³³⁵, fru>TNT, tsh-GAL80: n = 28). ^∗∗∗p < 0.0001, ^∗∗p = 0.0041 by Mann–Whitney test.