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

Fig. 4. Sequence of N protein involved in NLRP3 inflammasome activation.

Fig. 4

a Schematic diagram of wild-type SARS-CoV-2-N protein and truncated mutants N protein (N1 to N8). HEK293T cells (b) or A549 cells (d) were co-transfected with HA-NLRP3 and Flag-Ctrl, Flag-N truncated mutants (N1–N8). Cell lysates were immunoprecipitated using anti-Flag antibody and analyzed using anti-Flag and anti-HA antibody. Cell lysates (40 μg) were used as Input. c HEK293T cells were co-transfected with HA-NLRP3 and Flag-N truncated mutants (N1–N8). Cell lysates were immunoprecipitated using anti-HA antibody, IgG antibody was used as negative control, and analyzed using anti-Flag and anti-HA antibody. Cell lysates (40 μg) was used as Input. e HEK293T cells were co-transfected with plasmids encoding NLRP3, ASC, pro-Casp1, and pro-IL-1β, and transfected with plasmids encoding SARS-CoV-2-N protein and truncated mutants N protein (N1–N8) for 48 h. Supernatants were analyzed by ELISA for IL-1β and by WB for p17 and p20. Flag-ctrl means pcDNA3.1(+)–3× flag empty plasmid. Mock means untreated cells (e). Control means transfected empty plasmid (e). Data are representative of three independent experiments and one representative is shown. Error bars indicate SD of technical triplicates. Values are mean ± SEM. *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, two-tailed Student’s t-test. Source data are provided as a Source Data file.