The Interaction of Selectins and PSGL-1 as a Key Component in Thrombus Formation and Cancer Progression

Figure 7

During functional testing of E-selectin on M35/01 melanoma cells by CLSM, these cells were incubated with normal PBMCs for at least 4 hours in the absence (A) and presence of anti-PSGL-1 (B) (clone KPL-1) or anti-E-selectin (C) (clone P2H3) blocking antibodies (BD). Tumor cells were stained with anti-CD146-FITC (green), while anti-CD45-PE (red) (BD) was used for leukocytes to detect heterotypic cellular aggregates. A significantly decreased binding of PBMCs to cancer cells was seen with both blocking antibodies (B, C) suggesting the development of cell-cell interactions via E-selectin and PSGL-1 (A). Representative photos of all conditions are demonstrated. Flow cytometry analysis showed a similar tendency in the ratio of bound PBMCs to cancer cells with (B, C) or without blocking antibodies (A). Double positive events are expressed in % in the dot plots.