Fig. 7. Cell surface clusters. (A) Cells were transfected with myc-p25SS, incubated without permeabilization at 4°C with the ASSP mutant of aerolysin. Then, cells were treated simultaneously with chicken anti-aerolysin antibodies, Cy3-anti-myc antibodies (color-coded in green) and Cy5-transferrin (color-coded in red), to avoid possible accessibility problems due to raft clustering. Finally, raft domains were further clustered and visualized using anti-chicken secondary antibodies (color-coded in blue). An analysis by triple-channel fluorescence shows the distribution of each marker in a transfected cell, and the inset shows a high magnification view of the outlined area. (B) As in A, except that cells were transfected with p23-CD4TM, and transferrin was omitted. A high magnification view of the plasma membrane shows the distribution of ASSP, p23-CD4TM and the merged image (left). (C) Cells transfected with p25SS were processed for electron microscopy. The micrograph shows a high magnification view of the plasma membrane with immunogold labeling of p25SS. Scale bar: 2 µm.

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