Fig. 4: Western blot analysis of cytosolic and mitochondrial protein fractions of mosquito thoraxes indicating Coeae6g sub-cellular localization.

ATP5A and alpha-tubulin are used as mitochondrial and cytosolic markers, respectively. Approximately 30 μg of total protein deriving from the same experiment were loaded from each fraction, in two separates, simultaneously processed, 10% SDS gels, and immuno-blotted against the three protein targets; a single nitrocellulose membrane was stained initially against Coeae6g and, after mild stripping using a low pH glycine solution and re-blocking, it was re-blotted against alpha-tubulin. Mitochondria isolation experiments and subsequent western blot analyses were independently repeated twice, yielding similar findings.