Fig. 2

poly(dA:dT) reduces the amplified luminescent proximity signal between the PYD domain of AIM2 and the HIN-200 domain of AIM2. A schematic representation of full-length AIM2 (AIM2-FL) and its truncated proteins, the HIN200 domain of AIM2 (AIM2-HIN200) and PYD domain of AIM2 (AIM2-PYD). We synthesized two truncated forms of AIM2, AIM2-PYD-FLAG and AIM-HIN-200-Biotin, using a wheat germ cell-free synthesis system (a). Synthetic protein–protein interactions were detected by the amplified luminescent proximity homogeneous assay (Alpha). A total of 100 ng of each protein indicated was incubated with 5 μg/mL anti-FLAG mAb M2, 16.67 μg/mL protein-A-conjugated Alpha acceptor beads (PerkinElmer, Waltham, MA, USA), and 16.67 μg/mL streptavidin-conjugated Alpha donor beads (PerkinElmer, Waltham, MA, USA) for 24 h with or without 5 mg/mL poly(dA:dT) (Invivogen, San Diego, CA, USA). Responses (counts) were measured using EnSpire™ Multimode Plate Reader (PerkinElmer, Waltham, MA, USA). The results are given as means ± standard deviation from triplicate wells. Asterisk indicates significance (p < 0.01) (b)