Fig. 5

Colocalization of p27 and CD59 in unstimulated but not in activated lymphocytes. (A) Overlapping of p27 and CD59 expression in Triton X-100 treated lymphocytes. Cells treated with Triton X-100 were stained directly with (PE)-conjugated anti-CD59 and indirectly with anti-p27 as described in the text. Color channels were set so that CD59 appears green, p27 appears red, and overlap appears yellow. (B) Lack of aggregation of p27 in lymphocytes activated by CD59 crosslinking. Nonadherent peripheral blood mononuclear cells (PBMC) were activated with murine anti-CD59, crosslinking with anti-mouse immunoglobulin G-(IgG-FITC) and incubation at 37°C. They subsequently were stained indirectly with p27 as described. p27 staining (red) and with CD59 staining (green) is shown for three representative cells. p27 staining (red) of the cells is shown in the left column. The right column shows the same cells, gated to reveal both p27 (red) and CD59 (green) staining. Control staining is shown below, performed as above, but using an isotype control IgG instead of anti-p27. Staining with isotype control for CD59 (not shown) was negative.