Transcriptome analysis

Midguts of mosquitoes fed on S and G sugar meals were used for transcriptome analysis. Four mosquito midguts were pooled for one sample, and three biological replicates were collected from each treatment. Total RNA was extracted using TRIzol® Reagent according the manufacturer’s instructions (Thermo Fisher, China) and sent to Majorbio (Shanghai, China) for library construction and sequencing using Illumina HiSeq xten. Clean data were aligned to the reference genome AsteS1.7 (https://vectorbase.org/organisms/anopheles-stephensi). The expression level of each transcript was calculated according to the fragments per kilobase of exon per million mapped reads (FRKM) method. R statistical package software EdgeR (Empirical analysis of Digital Gene Expression in R, http://www.bioconductor.org/packages/2.12/bioc/html/edgeR.html) was utilized for differential expression analysis.