Identification of CD226 ligand on colo205 cell surface

Figure 1 Sequence of CD226 cDNA and primers. 174-923 bp: extracellular region of CD226; 174-227 bp: signal peptide sequence; 924-998 bp: transmembrane region; labeled sequence: primers; under-lined sequence was the splicing donor sequence.

Figure 2 Construction of pCD226/Ig fusion vector.

Figure 3 Identification of CD226/Ig fusion protein purified by Leo-A1 Sepharose-4B affinity column. 1. Supernatant of pIG vector transfected COS-7 cells; 2. Supernatant of pCD226/Ig transfected COS-7 cells; 3. Supernatant of pCD226/Ig trans-fected COS-7 cells after concentrated by ammonium sulfate; 4. Flow through solution of pCD226/Ig transfected COS-7 cells after affinity chromatography; 5. CD226/Ig fusion protein purified by Leo-A1 sepharose-4B affinity column; 6. Marker.

Figure 4 Western-blot result of CD226/Ig by anti-IgG polyclonal antibody Lane 1. Supernatant of pCD226/Ig transfected COS-7 cells; Lane 2. Super-natant of pIG transfected COS-7 cells

Figure 5 CD226L expression on Colo205 cells identified by CD226/Ig immunohistochemistry. A: Colo205 negative control (stained with hIg); B: Colo205 cells were specifically stained by CD226/Ig.

Figure 6 Adhesion experiments of activated Jurkat cells with Colo205 cells. Density of Colo205 was 0.5 × 10⁵ per well. Density of Jurkat was 1 × 10⁵ per well. R stand for resting Jurkat cells. A stand for activated Jurkat cells.