FIGURE 4.

Effects of DAP12 on TREM2-CTF require their interaction. A, HEK293T cells were co-transfected with TREM2-Myc and wild-type (DAP12 WT-GFP) or mutant (DAP12 D50A-GFP) DAP12 plasmids. Lysates were immunoprecipitated using antibodies against mouse IgG (M IgG), mouse anti-Myc antibody (Myc), rabbit IgG (R IgG), or rabbit anti-GFP antibody (GFP), followed by immunoblotting with antibody against the Myc or GFP epitope. Five percentage of total protein was loaded as an input control. Arrowheads indicate each specific band recognized by the antibodies or the IgG heavy or light chain. B, HEK293T cells were co-transfected with TREM2-Myc and wild-type (DAP12 WT-GFP) or mutant (DAP12 D50A-GFP) DAP12 plasmids. Cells were harvested 24 h after transfection, and the lysates were analyzed by Western blotting. Bar graph to the right shows the quantification of Western blots as ratios of TREM2-CTF/TREM2-FL (n = 3). **, p < 0.01.