ER proteins listed in Table 1 and Table S1were RNAi-mediated knockdown of alpha-COPI expression and validation by QRTPCR alpha-COPI gene was subjected to a RNAi according to our previous study [31] . Briefly, a DNA fragment of alpha-COPI gene was amplified by PCR (forward primer containing the T7 promoter sequence: 59 TAATACGACTCACTATAGGGAGA TGCTGACAAATTTCGAAACCAA 39; and reverse primers containing the T7 promoter sequence: 59 TAATACGACTCAC TATAGGGAGATCCGTCGCCGTAGGATTCTT 39) and cloned into the pGEM-T easy vector (Promega). Subsequently, a double-strand RNA (dsRNA) was synthesized in vitro transcription using the MEGAscript RNAi Kit (Ambion). Two-day old female mosquitoes were injected with 400 ng of dsRNA using a Nanoject II microinjector (Drummond Scientific). The knockdown efficiency of mRNA encoding alpha-COPI was determined using mosquitoes injected with Fluc (firefly luciferase) dsRNA as a control. QRT-PCR was performed using PerfeCTa SYBR Green FastMix (Quanta BioSciences) by Real-Time PCR (7300 Real-Time PCR System, Applied Biosystems) with alpha-COPI forward primer: 59 GTGTCCGCATCGTTGGATCA 39, alpha-COPI reverse primer: 59 ACAACAGCATCAGCTTGCCCAA 39. Ribosomal protein S7 mRNA levels were used as an internal control for normalization. Statistical analysis of the gene expression was done by unpaired student t test using GraphPad Prism software (GraphPad Software, Inc.).