Hemolymph Collection.

Hemolymph of adult female mosquitoes was obtained by perfusion as previously described (65). Fine needles were made from 100-μL glass microcapillary tubes using a micropipette puller P-30 (Sutter Instrument), and mounted on a pump (Drummond). Needles were inserted manually through the neck membrane into the thoracic cavity, and insects were perfused with 20 μL of phosphate buffered saline (PBS, pH 7.2). The hemolymph was obtained from a small tear made laterally on the intersegmental membrane of the last abdominal segment. The first drop of perfused hemolymph was collected into a silanized glass tube (Thermo Fisher) placed on ice.