Sensory treatment
We first exposed 3 day-old wildtype (wt ) virgin males to females
cues for 24 hours. To do so, we isolated standard males in a vial, and
this vial was connected to either a) another vial containing three 3
day-old virgin wt females (i.e. female-exposed male) or b) to an
empty vial (control male). Importantly, interconnected vials were
separated by a fine mesh partition, and this allowed exchange of female
semio-chemicals (volatiles but also probably non-volatiles) as well as
female visual cues across the chambers, while ensuring males would not
mate (GarcĂa-Roa, Serra and Carazo, 2018). Previous empirical research
has determined that this methodology does not elicit any courtship
behaviour in female-exposed males (Corbel et al. , 2022).