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. Author manuscript; available in PMC: 2019 Oct 5.
Published in final edited form as: Science. 2019 Mar 14;364(6435):eaau1330. doi: 10.1126/science.aau1330

Fig. 2. Degradation of NLRP1B is necessary and sufficient for NLRP1B inflammasome activation.

Fig. 2.

(A) 293T cells transfected with constructs encoding NLRP1B, CASP1 and pro-IL-1β were assayed for inflammasome activation as in Fig. 1 in the presence (+) or absence (−) of proteasome inhibitors (MG132 (10 μM) and Bortezomib (1 μM)) or p97/VCP inhibitor (NMS-873 (0.5 μM)). (B) Immortalized 129 bone-marrow macrophages were treated with lethal factor (LF) to activate the NLRP1B inflammasome ± MG132. Endogenous NLRP1B was detected by immunoblot (IB) with 2A12. Relative band intensities are indicated. (C) The plant auxin-interacting degron (AID) was fused to the N-terminus of indicated GFP-NLRP1B variants. Specific degradation was induced with indole-3-acetic acid (IAA) in TIR1-expressing 293T cells. Inflammasome activation was assessed by immunoblot (IB) for IL-1β p17. Relative band intensities are indicated. Gel images are representative of experiments performed at least three times. S, serine. A, alanine.