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. 2021 Aug 2;12:4664. doi: 10.1038/s41467-021-25015-6

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© The Author(s) 2021, corrected publication 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. 6 — a HEK293T cells (top) or A549 cells (bottom) were transfected with Flag-NC/HA-NLRP3, Flag-N/HA-NLRP3, or Flag-3a/HA-NLRP3 for 24 h. The sub-cellular locations of NLRP3 (red), Nucleus marker DAPI (blue), and SARS-CoV-2 N or SARS-CoV-2 3a (green) were visualized with confocal microscopy. b HEK293T cells (top) or A549 cells (bottom) were transfected with Flag-N2+HA-NC or Flag-N2+HA-NLRP3 for 24 h. The sub-cellular locations of NLRP3 (red), Nucleus marker DAPI (blue), and SARS-CoV-2-N8 (green) were visualized with confocal microscopy. c HEK293T cells (top) or A549 cells (bottom) were transfected with Flag-N8+HA-NC or Flag-N8+HA-NLRP3 for 24 h. The sub-cellular locations of NLRP3 (red), Nucleus marker DAPI (blue), and SARS-CoV-2-N8 (green) were visualized with confocal microscopy. d, e PMA-differentiated THP-1 macrophages were stably infected with Lentivirus-CT or Lentivirus-N. The sub-cellular locations of SARS-CoV-2-N (green, d) Nucleus marker DAPI (blue), and NLRP3 (red, e) were visualized with confocal microscopy. f THP-1 macrophages were stably infected with Lentivirus-CT or Lentivirus-N, and stimulated by LPS (1 μg/ml) plus ATP (2.5 mM) or LPS (1 μg/ml) plus Nigericin (2 μM). ASC oligomerization was analyzed by immunoblotting. g GM-CSF differentiated mice BMDMs were infected with Lentivirus-CT or Lentivirus-N and stimulated with LPS (1 μg/ml), LPS (1 μg/ml) plus ATP (2.5 mM), or LPS (1 μg/ml) plus Nigericin (2 μM). ASC oligomerization was analyzed by immunoblotting. h HEK293T cells were co-transfected with plasmids encoding NLRP3, ASC, pro-Caspase-1, or pro-IL-1β, and transfected with plasmids encoding SARS-CoV-2-N protein and truncated mutants N protein (N1 to N8) for 48 h, ASC oligomerization was analyzed by immunoblotting. Flag-NC or HA-NC means pcDNA3.1(+)–3× flag or pCAGGS-HA empty plasmid (a–c). Mock means untreated cells (f, h). Control means transfected empty plasmid (h). Data are representative of three independent experiments. Source Data are provided as a Source Data file.