FIG 6.

Examination of ORF2, β-catenin, and HMGA1 in transiently transfected Neuro-2A cells. Neuro-2A cells were cotransfected with 0.4 μg of each plasmid expressing β-catenin (non-Flag-tagged protein), HMGA1, and Flag-tagged ORF2 by use of the Lipofectamine 2000 transfection reagent (Invitrogen). At 48 h posttransfection, cells were prepared for confocal microscopy. (A) Neuro-2A cells were stained with primary antibodies against β-catenin (green) and Flag to detect ORF2 (red). (B) Neuro-2A cells were stained with a primary antibody directed against HMGA1 (green) or Flag to detect ORF2 (red). Nuclear DNA was stained with DAPI. Images were observed by confocal microscopy, and these images are representative of those from four experiments. Bars, 10 μm.