Midgut peroxides and epithelial integrity were altered at 72-IGF1 feeding.

<p>Peroxide levels in midguts at 72(A) and one week (D) following IGF1 treatment. Peroxides were quantified in pools of five midguts; three pools were collected per treatment per experiment. Experiments were replicated three times with separate cohorts of mosquitoes. Graphs show the mean fold induction of peroxides±SEM in IGF1 treated groups relative to saline/ATP-fed controls. Statistical significance was determined by unpaired t-test and p-values are indicated. Midgut caspase-3 activity at 72 h (B), one week (E), two weeks (F), and three weeks (G) following IGF1 treatment. Graphs show the average fold induction±SEM of AFC fluorescence (caspase-3 activity) relative to controls. Three pools of five midguts per treatment were collected for 72 h and one week timepoints and 1–2 pools were collected for two and three week timepoints. Experiments were replicated 4–6 times with separate cohorts of mosquitoes. Caspase-3 activity among groups was compared using Kolmogorov-Smirnov test. (C) Midgut permeability at 72 h following a blood-meal containing IGF1 and 3–3.5 µm fluorescent beads (Spherotech). Each data point represents the number of fluorescent beads in three whole mosquitoes quantified by flow-cytometry. Statistical significance was determined by one-way ANOVA followed by Fisher's LSD.