Subcellular localization of APOBEC proteins. HeLa cells were transfected with 5 μg each of pcDNA-Apo3G (panel A), pcDNA-A3A (panel B), or pcDNA-A3G-3A (panel C). Immediately following transfection, cells were detached from the flasks by trypsinization and re-seeded into 12 well plates containing microscope cover slips. Transfected cells were grown on the cover slips over night and then fixed with ice cold methanol for 10 minutes (-20°C). Cells were then stained with an A3G-specific rabbit polyclonal antibody (ApoC17) and analyzed by confocal microscopy as detailed in Materials and Methods.