Journal: Journal of Virology
Article Title: Effects of Manipulating Apoptosis on Sindbis Virus Infection of Aedes aegypti Mosquitoes
Figure Lengend Snippet: Caspase activity in midgut and remaining carcass following injection of dsRNA. Rexville D mosquitoes were injected with dsRNA ( Aeiap1 , Aedronc , or cat ) or PBS or mock injected. Midguts were dissected at 1, 2, or 3 days postinjection, and pooled tissues from 10 individuals were homogenized in lysis buffer. Caspase activity was determined in midguts (A) and remaining carcass (B) using the effector caspase substrate Ac-DEVD-AFC. Data are means ± standard errors of the means from three independent experiments, and statistical significance (****, P
Article Snippet: Protein concentration was determined by absorbance at 280 nm, and 50 μg of protein was mixed in 100 μl reaction buffer (100 mM HEPES [pH 7.4] containing 2 mM DTT, 0.1% 3-[(3-cholamidopropyl)-dimethylammonio]-1-propanesulfonate [CHAPS], 1% sucrose) with 200 μM Ac-DEVD-AFC (MP Biomedical), an effector caspase substrate, and incubated for 15 min at 37°C.
Techniques: Activity Assay, Injection, Lysis