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Fig. 4. Concentration of HTLV-1 envelope glycoproteins and hDlg in restricted areas characteristic of cell contact sites in HTLV-1-infected primary T lymphocytes. Each row shows a single optical plane of HTLV-1 infected CIB cells, as recorded independently in the red and green channels by confocal microscopy. (A,B) HTLV-1 envelope glycoproteins were stained using sera obtained from infected patients, and Texas-red conjugated secondary antibodies before permeabilization (left panels). hDlg (A) and Gag (B) were stained on saponin-permeabilized cells using 2D11 and p19 monoclonal antibodies, respectively, followed by FITC-conjugated secondary antibodies (middle panels). On the right panels are presented overlay profiles of red and green channels (Overlay panels). Co-localization signal appears as yellow pixels. (B) HTLV-1 envelope glycoproteins were stained as described above (left panels). Staining of CD25, CD2 and CD4 were performed on non-permeabilized cells using either FITC-conjugated mAb, or mAb followed by FITC-conjugated secondary antibodies (middle panels). Staining of GM1 was done on non-permeabilized cells with biotin-conjugated cholera toxin and DTAF-conjugated streptavidin. Staining of Lck was performed on saponin-permeabilized cells using rabbit sera followed by FITC-conjugated secondary antibodies. Overlay profiles of red and green channels. Co-localization signal appears as yellow pixels (right panels, merge).

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