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. 2021 Jan 8;12:188. doi: 10.1038/s41467-020-20319-5

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© The Author(s) 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 3 — a Top panel: SDS-PAGE NLRP1-FIIND^UPA-CARD mutants. Bottom panel: native gel analysis of SUMO-NLRP1-FIIND^UPA constructs, stained by Krypton. b ASC-GFP speck formation induced by wild-type NLRP1-FIIND^UPA-CARD and PP1278AG in 293T-ASC-GFP cells. c Percentage of ASC-GFP specks induced by NLRP1-FIIND^UPA-CARD constructs in 293T-ASC-GFP cells. Cells were fixed 24 hours after transfection for fluorescence microscopy. Data are presented as mean values ± SD. P value was calculated with one-way ANOVA, n = 3 biological replicates. ‘*/**/****’ indicates P value < 0.05, 0.01, and 0.0001, correspondingly. d Side view and top view of FIIND^UPA-dimer structures reported by other groups, and our modeled oligomer interface of NLRP1-FIIND^UPA. Potential surface residues that may involve homotypic oligomerization were displayed using spheres, PP1278-1279, were highlighted in red color, R1240, RK1260, R12785, and F1320 are colored in orange. e The top and side views of NLRP1-CARD filament and six modeled FIIND^UPA domains fitted into the FIIND^UPA-CARD filament density. f A close-up view of the potential UPA-CARD linker orientation. A linker with 25~40 Å is likely to support a regular organization of the two-layer structure. g The top and side views of CARD8-CARD filament and si modeled FIIND^UPA domains fitted into the CARD8-FIIND^UPA-CARD filament density.