CD84 and Ly-9 colocalize with SAP in Jurkat after activation of the receptors with monoclonal antibody.

A fluorescence microscopy approach was used in the human T-cell line Jurkat to confirm the SAP/Ly-9 and SAP/CD84 interaction in T cells. Jurkat cells were incubated with 5 μg/mL biotinylated anti-CD84 or anti-Ly9 antibody for 30 minutes at 4°C, as detailed in “Materials and methods.” Cells were then incubated with streptavidin-FITC and incubated at 4°C (control) or 10 minutes at 37°C (capping). Cells were then stained with Cy3-SAP, as indicated. Green fluorescence shows CD84 (A, C) or Ly 9 (E, G). Staining with red fluorescence indicates SAP (B, D, F, H). White arrows show colocalization.