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Activation of Akt Signaling Reduces the Prevalence and Intensity of Malaria Parasite Infection and Lifespan in Anopheles stephensi Mosquitoes

Figure 3

Membrane localization of the transgene.

A. Immunocytochemistry of myr-AsteAkt-HA transgenic or non-transgenic paraffin embedded midgut sections using an anti-HA-fluorescein antibody. Images were acquired under brightfield illumination to visualize the midgut epithelial architecture or a FITC filter to visualize the overexpressed protein. To determine localization the images were merged. Arrows indicate the cell membrane of the midgut epithelial cells. Five midguts from transgenic and non-transgenic mosquitoes were analyzed. B. Immunocytochemistry of myr-AsteAkt-HA transgenic or non-transgenic midgut sheets using an anti-HA-fluorescein antibody. At least 10 midgut sheets from three separate cohorts of mosquitoes were analyzed by confocal microscopy. Three representative myr-AsteAkt-HA transgenic and non-transgenic midguts are shown. C. Membrane (M), cytoplasmic (C), and nuclear (N) fractions of transgenic mosquito midguts were prepared and these fractions, and an intact midgut sample (MG), were probed with anti-HA antibody. This immunoblot is representative of three experiments.

Figure 3