Newly hatched to one day-old flies were collected and transferred to
fresh food vials. The following day at 17:00 hrs, 20 flies of
mixed sex were either transferred into starvation vials (see
Starvation procedure) or kept in the food vials. After 20 h,
approximately 40 female flies per group were cold-anesthetized,
pierced through the thorax by the tip of a dissecting needle (0.5 mm
in diameter), and collected on ice within a sieve composed of two
tubes. The hemolymph was centrifuged out of the fly into the bottom
tube at 4°C. 0.5 µl of the extracted hemolymph was transferred
by a capillary (0.5 µl, Hirschmann Laborgeraete, Eberstadt,
Germany) into 19.5 µl PBS (see dx.doi.org/10.17504/protocols.io.dkn4vd).