Fig. 4. Mitochondrial and cytosolic protein acetylation in Aedes aegypti fat body and thorax.

Newly-eclosed mosquitoes were maintained on 3% sucrose until day 4 post-eclosion, and then fed on either 3% sucrose or water for 72 h. Mitochondrial and cytosolic protein fractions were isolated from the fat body (A) and thorax (B) of mosquitoes fed on either 3% sucrose or water for 72 h. Western blotting was performed using an anti-acetyl lysine antibody. The acetylated protein bands detected by western blots were quantified and normalized to porin or α-tubulin (C and D). Densitometric analysis was performed with NIH Image J software (Schneider et al., 2012). Each lane contains 20 μg of proteins. Anti-porin and anti-α-tubulin antibodies were used as loading controls for mitochondrial and cytosolic proteins, respectively. Western blots are representative of 3 independent biological replicates. Each protein extract replicate was prepared from a pool of approximately 200 mg of tissues. For fat body tissues isolated from sucrose- and water-fed mosquitoes, N = 1,216 (n = 333, 444, and 439) and 2,143 (n = 746, 702, and 695), respectively. For thorax dissected from sucrose- and water-fed mosquitoes, N = 543 (n = 169, 201, and 173) and 579 (n = 233, 192, and 154), respectively. Data represent the mean fold change ± SEM of 3 independent western blots. An unpaired Student’s t-test was employed. No significant difference (NS) was observed between treatments. M, protein markers; S, sucrose; W, water.