For triglyceride and glycogen, a single fly was anesthetized and transferred to tube with the corresponding extract. The samples were measured according to the manufacturer’s instructions. Glycogen was measured with Liver/Muscle glycogen assay kit (#A043-1-1, Nanjing Jiancheng Bioengineering Institute, China). Triglyceride was measured with Triglyceride Quantification Colorimetric/Fluorometric Kit (#A110-1-1, Nanjing Jiancheng Bioengineering Institute).
The hemolymph trehalose was quantified by trehalose quantification kit (#A149-1-1, Nanjing Jiancheng Bioengineering Institute). Briefly, 40 flies were anesthetized and then pierced in the thorax with dissecting forceps. The pierced flies were then transferred to perforated tubes and centrifuged for 5 min at 3000×g at 4°C to collect the hemolymph. Afterward, 0.6 μL hemolymph was quickly removed into a 200 μL tube with trehalose extract and vortexed for 2–3 min. After 45 min standing, the sample was centrifuged at 8000×g for 10 min. Then 175 μL supernatant was added to 700 μL reaction solution and boiled for 5 min. After cooling, 250 μL sample was removed to a 96-well plate for the light absorption value measurement at 620 nm.
The hemolymph trehalose was quantified by trehalose quantification kit (#A149-1-1, Nanjing Jiancheng Bioengineering Institute). Briefly, 40 flies were anesthetized and then pierced in the thorax with dissecting forceps. The pierced flies were then transferred to perforated tubes and centrifuged for 5 min at 3000×g at 4°C to collect the hemolymph. Afterward, 0.6 μL hemolymph was quickly removed into a 200 μL tube with trehalose extract and vortexed for 2–3 min. After 45 min standing, the sample was centrifuged at 8000×g for 10 min. Then 175 μL supernatant was added to 700 μL reaction solution and boiled for 5 min. After cooling, 250 μL sample was removed to a 96-well plate for the light absorption value measurement at 620 nm.