Activation of Akt Signaling Reduces the Prevalence and Intensity of Malaria Parasite Infection and Lifespan in Anopheles stephensi Mosquitoes
A. Total RNA was isolated from the midguts and carcasses of ten non-bloodfed (NBF) and ten bloodfed transgenic mosquitoes at 2 h, 24 h, 48 h and 72 h post-bloodfeeding and converted into cDNA. The 72 h sample was collected post-oviposition. Transgene specific qRT-PCR primers were used to amplify myr-AsteAkt from the cDNA; amplification of ribosomal protein S7 was used for normalization. The qRT-PCR experiments were performed in triplicate and replicated twice with separate cohorts of mosquitoes. Data are represented as means ± SEMs. B. Total protein was isolated from the midguts of transgenic mosquitoes at various time points during a reproductive cycle. Proteins were blotted and probed with anti-HA antibody or anti-GAPDH antibody to assess protein loading. C. Average expression of transgenic protein normalized to GAPDH loading controls and shown relative to levels in non-bloodfed mosquitoes (NBF). Data are represented as means ± SEMs from four replicates with separate cohorts of mosquitoes.