Extended Data Fig. 2: Further characterization of integrative properties of multimodal threats during mating.

(a) Flies were placed in elevated behavioral arenas connected to two-way solenoids. Compressed air sources were fed into airflow meters and then into the solenoids, which gate the delivery of the wind. By controlling solenoid opening via Arduino, specific gusts of wind of timed duration can be delivered to behavioral arenas. A camera was suspended above the behavioral arenas. For more information, see Methods. (b) Side view of each well. Tubes were connected into wells via adapters, and a mesh layer was placed over the hole in the floor so the flies did not fall in. (c) Photo of the setup. Solenoids are controlled via a computer connected to the Arduino (not in view). (d) Flies end matings in response to a 350-millisecond wind gust with varying latencies. Cumulative distribution plot of the 10- and 15-minute data in Fig. 2d. (e) Single 650-millisecond wind gusts at 10 min into mating, and 250-millisecond wind gusts at 15 min into mating each terminate ~30% of matings. Data used to calculate independent probability of paired pulse experiment in Fig. 2f. (f) Of 164 descending-interneuron-labeling lines screened, none terminated mating at 5 min when stimulated for 15 seconds with CsChrimson. SS01593 (containing the serotonergic descending neuron DNg26, as well as labeling other cells) is the most effective line at terminating mating at 15 min. (g) DNg26 (SS01593) sends projections to the abdominal ganglion. (h) 700 milliseconds of DNg26 stimulation at 10 min into mating, or 300 milliseconds of DNg26 stimulation at 15 min into mating terminate ~30% of matings. (i) Paired pulse stimulation (700 ms at 10 min, 300 ms at 15 min) of DNg26 is integrated over a longer timescale when delivered at 15 min into mating.

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