Fecundity experiment
We measured egg lying rate, fertility, fecundity, and egg to adult
survival of females mated with a temperature challenged male that was
either allowed to recover or not. Mating assays took place on three
different days: two, four and six days after adult male eclosion to
assess changes in fertility during sexual maturation. Males were paired
with five-day old virgin females in all cases. New virgin males were
used on each mating day and 40 pairs per treatment and day were
initially set up. Pairs were allowed to mate for seven hours on day two
and four, and for five hours on day six. The mating was observed, and
the duration was adjusted each day in order to have an appropriate
sample size for all treatments (see Supplementary table S1 for the full
sample sizes). At the end of the interaction time, males were discarded.
Mated females were kept at 25°C until the next day allowing them to lay
eggs and were then discarded. We counted the number of eggs and the
vacated vials were kept at standard conditions for 12 days allowing all
the offspring to eclose. After that time, the number of offspring was
counted, and we estimated the egg to adult survival. This assay was
repeated independently following the same procedure and adding one
mating day (one, two, four and six days after adult male eclosion). With
that, we could get a better understanding of the effects of heat stress
on fertility and fecundity in recently eclosed males.