Extended Data Fig. 4: Toxins mediated Rho GTPases activation but not inhibition trigger the NLRP3 inflammasome.
a, BMDMs isolated from wild-type or NLRP3 knock out C57BL/6 J mice were treated with DNT (1 µg/mL) for 8 h. Supernatants and cell lysates were analyzed by immunoblot. b-c, HEK293T cells were transfected as indicated with plasmids encoding NLRP3 inflammasome components (myc-NLRP3, ASC-GFP, mCaspase-1) and pro-IL-1β-Flag together with (b) myc-DNT, HA-YopE or myc-Dbl495–826 or (c) transfected with SopE-HA and treated with MG132 to block SopE degradation (10 μM). Supernatants and cell lysates were analyzed by immunoblot. d, BMDMs isolated from C57BL/6 J mice were treated with IPA-3 (5 μM) or MCC950 (1 μM) for 45 min prior to 8 h of DNT treatment (1 μg/mL). Supernatants and cell lysates were analyzed by immunoblot. The numbers on the side of the immunoblots indicate molecular weight (kDa). Experiments were repeated at least three times, and representative data are shown.
