CAFE assay

We modified the CAFE assay⁴² by putting three flies in a transparent vial containing 1cm high 1% agar at the bottom to keep the moisture. Two 5ul capillaries were inserted into a cotton plug on top. One capillary contained liquid food, including 10% yeast, 10% sucrose, and blue dye, while the other one contained liquid food mixed with Ecc15. To make Ecc15-mixed liquid food, 40ml Ecc15 was freshly cultured as described above, and was resuspended in 3ml liquid food. The food intakes were recorded at corresponding time points and the capillaries were changed every 24 hours.