Figure 3.
Quantification of TREM2·TYROBP complex by “sandwich” ELISA. HEK293 cells were transfected with TREM2-CLuc-IRES-TYROBP-NLuc followed by anti-TREM2 antibody (Ab) stimulation to induce TREM2 coupling to TYROBP and protein cross-linking with DTBP and phosphatase inhibitor sodium pervanadate (Na3O4V) (A). Following cell lysis, protein was incubated in anti-TYROBP-coated wells and labeled for TREM2 with biotinylated anti-TREM2·HRP-conjugated streptavidin complex for quantification of TREM2·TYROBP protein complexes. Average (Ave) OD was quantified by ELISA. Data represent the arithmetic difference between a given treatment condition and the average OD of untransfected controls (n = 4 per group) (B). Significant differences were determined by one-way ANOVA followed by Tukey's post-test. Error bars represent S.D. †† and ††† denote p < 0.01 and p < 0.001, respectively, versus untransfected cells. ### denotes p < 0.001 versus unstimulated cells transfected with TREM2-TYROBP. ** and *** denote p < 0.01 and 0.001, respectively, versus those indicated.