FIG 9.

Localization of Akt3 and ORF2 in transfected Neuro-2A cells. Neuro-2A cells were transfected (Lipofectamine 2000; Invitrogen) with 0.5 μg of a plasmid that expresses Akt3 (green) (A), Flag-tagged ORF2 (red) (B), or Akt3 and ORF2 (C). At 48 h posttransfection, cells were fixed and then stained with antibodies that recognize Akt3 (green-stained cells) (ab1521; Abcam) and/or ORF2 (red-stained cells). Nuclear DNA was stained with DAPI. Images were observed by confocal microscopy. The selected images are representative of 4 experiments (at least 100 stained cells were examined). For panel C, we show ORF2 alone, Akt3 only, and merged images that also show DAPI-stained cells that are ORF2 and Akt3 positive. The scale bar in each panel is equal to 50 μm.