Figure 4. In vitro reconstitution of the AIM2 inflammasome.

a, Immunoblot for caspase-1 (upper panel) or IL-1β (lower panel) following in vitro incubation of PYD-caspase-1 and pro-IL-1β with purified GST-AIM2 (2^nd lane), GST-AIM2-ΔPYD (3^rd lane) or ASC pyroptosome (4^th lane). b, c, Autoradiograms of ³⁵S-labeled WT or C287A chimeric PYD-caspase-9 following in vitro incubation with purified GST-AIM2 (2^nd and 6^th lanes), GST-AIM2-ΔPYD (3^rd and 7^th lanes) or ASC pyroptosome (4^th and 8^th lanes) (b) or with the indicated amounts of purified GST-AIM2 in the presence or absence of the indicated amounts of poly (dA:dT) (c). d, Immunoblot for AIM2 after cross-linking of purified AIM2 (2 µg) with glutaraldehyde in the presence of the indicated amounts of 64-mer dsDNA. A longer exposure of this gel is shown in Supplementary Fig. 8.