ice-cold 1.5-ml Eppendorf tube containing 100 ml of 16Isotonic Extraction Buffer (5 mM HEPES;pH 7.8, 0.25 M sucrose, 1 mM EGTA, 25 mM KCl), and an appropriate amount of Protease Inhibitor Cocktail. Mosquito midgut ER protein extraction was performed according using the Endoplasmic Reticulum Isolation kit (ER0100; Sigma, St. Louis, USA) with modification. Briefly, the dissected midguts were homogenized with a blue Kontes pestle and spun at 1000g for 10 min at 4uC. The post nuclear supernatant was transferred into a clean 1.5-ml tube and spun at 12000g for 15 min at 4uC. The post mitochondrial supernatant was transferred into a clean 1.5-ml tube and mixed with 7.5 volumes of 8 mM CaCl 2 by vortexing. The mixture was incubated on ice for 10 min and then spun at 8,000g for 10 minutes at 4uC. The supernatant was discarded, and the pellet containing the microsomal fraction, was resuspended in 20 ml of 16 Isotonic Extraction Buffer followed by a 10-min of incubation on ice. A 5 ml portion of the extracted ER protein fraction was used for measuring protein concentration with the BCA TM Protein Assay Kit (Pierce, USA). The remaining ER proteins were mixed with an appropriate amount of 56SDS sample loading buffer, boiled for 4 min, and spun at 130006 rpm for 5 min. The supernatant was stored at 220C for SDS-PAGE.