Newly hatched to one day old flies were collected and transferred to fresh food vials. The following day, flies were briefly CO2-anesthetized and sorted by and genotype. At 17:00 hrs, around 20 female flies were transferred into a starvation vial (see Starvation procedure). Dead flies were counted every 3 h and not removed. Daily counting sessions were repeated from 9:00 to 18:00 hrs, until all flies were found dead.