Figure 2. The work flow from collecting flies to experimental assessments.

Prior to endurance training, a total of 1,060 male flies should be collected per genotype being assessed. Male flies are used because they exhibit a more robust exercise response than females. Flies should be divided into two groups, exercised and unexercised, with 530 flies in each group. After three weeks of endurance training, post-training assessments are performed during the fourth week (within one week after the last day of exercise). Lethal assessments are shown in black, then the assessment is lethal to the flies. It is possible to complete all of the experimental assessments with only 860 male flies per genotype, if flies from the longitudinal climbing assessment are also used for the fatigue assessment.