Figure 2

p21/waf1 and cyclin D2 colocalize in HTLV-1 infected cells. Cells were fixed with 2% paraformaldehyde and stained with rabbit polyclonal anti-cyclin D2 and mouse monoclonal anti-p21/waf1 antibodies, washed, and then stained with the secondary antibodies TR goat anti-mouse IgG and FITC goat anti-rabbit IgG. TOTO-3, a dimeric cyanine nucleic acid stain, was used as a nuclear stain. For induction of virus, TNF-α (10 ng/ml) was added for four hours. Confocal optical sections (z = 0.5 μm) are shown in all panels. In the nuclear panel, dark blue staining represents the nucleolus, whereas the lighter blue staining represents the nucleoplasm. The fourth column contains the merged FITC and TR channels. Arrows indicate points where colocalization is occurring, shown as yellow coloring when the two images are merged. Experiments were repeated three times and a representative sample from one experiment is shown.