Protein Extraction and Western Blotting Protocol

Protein extracts were prepared as previously described [24 (link),25 (link)]. Briefly, A. aegypti total protein extracts were carried out by homogenizing adult female mosquitoes or Aag2 cells in TBS containing a protease inhibitor cocktail (Sigma). Proteins were recovered from the supernatant by centrifugation at 14.000xg, for 15 min. at 4°C. Protein concentration was determined by the Bradford Protein Assay (Bio-Rad). Western blots were carried out using secondary antibody (Immunopure goat anti-mouse, #31430). The primary monoclonal antibodies (ChIP grade) used were anti-H3 pan acetylated (Sigma-Aldrich #06–599), anti-H3K9ac (Cell Signaling Technology #9649) and Anti-H3K27ac (Cell Signaling Technology, #8173), according to the manufacture’s instructions. For all antibodies, a 1:1000 dilution was used. For normalization of the signals across the samples, an anti-histone H3 antibody (Cell Signaling Technology, #14269) was used.