Figure 5: apterous neurons required for female receptivity are not part of the doublesex and fruitless circuitry.

(a) ap^GAL4/UAS-CD8-GFP female brain (3 days) stained with anti-GFP (green) and anti-Dsx (magenta). No coexpression between ap^GAL4 and Dsx was detected in the brain. Scale bar represents 50 μm except inset where it represents 15 μm. (b) Labeling of apterous and fruitless-positive neurons in the female brain (3–6 days) using ap^GAL4/UAS-DsRed (magenta) and fru^LexA/LexAop-CD2-GFP (green). Coexpression of ap^GAL4 and fru^LexA was detected in two neurons in the posterior brain (arrows). Scale bar represents 50 μm. (c) ap^GAL4/fru^LexA intersecting neurons labelled in females aged for 15 to18 days at room temperature. Intersecting neurons were visualized with anti-GFP (green) and nc82 (magenta). Scale bar represents 50 μm. (d) Quantification of mating of virgin females in which overlapping apterous and fruitless circuitry was silenced with a schematic representation of the manipulation employed depicted. Genotypes indicate virgin females. n values shown in parentheses. n.s., not significant, *p < 0.05, ***p < 0.001, Fisher’s exact test. (e) Quantification of mating of virgin females with ap^GAL4 neurons ‘masculinized’. Genotypes indicate virgin females. n values shown in parentheses. n.s., not significant, Fisher’s exact test.