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. 2023 Feb 16;14:872. doi: 10.1038/s41467-023-36522-z

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© The Author(s) 2023

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 Non-reducing immunoblotting of GSDMD oligomer in BM neutrophils isolated from WT, PIL mice and Gsdmd^−/− mice after pristane treatment. b Quantitative analysis of GSDMD oligomer/GAPDH. c Non-reducing immunoblotting of GSDMD in neutrophils from HV and SLE patients. d Quantitative analysis of GSDMD oligomer/GAPDH. n = 6 HVs or 6 SLE patients. e Non-reducing immunoblotting of GSDMD oligomer in murine BM neutrophils. Cells were treated with IFN-γ plus LS, or pretreated with Mito-TEMPO. f Quantitative analysis of GSDMD oligomer/GAPDH. g Immunoblotting of GSDMD in murine BM neutrophils. Cells were treated with IFN-γ plus LS, or pretreated with Mito-TEMPO. h Quantitative analysis of GSDMD-N/GAPDH. i Non-reducing immunoblotting of GSDMD oligomer in BM neutrophils isolated from MRL/mpj, MRL/lpr or MRL/lpr after Mito-TEMPO treatment. j Quantitative analysis of GSDMD oligomer/GAPDH. k Immunoblotting of GSDMD in BM neutrophils isolated from MRL/mpj, MRL/lpr and MRL/lpr after Mito-TEMPO treatment. l Quantitative analysis of GSDMD-N/GAPDH. m Non-reducing immunoblotting of GSDMD oligomer. Purified GSDMD protein was incubated with purified caspase-4 protein and LPS in vitro, then added with human mtDNA or Ox-mtDNA (20 nM). n Quantitative analysis of GSDMD oligomer/GSDMD monomer. o Non-reducing immunoblotting of GSDMD oligomer. The LPS/caspase-4/GSDMD system was added with Ox-mtDNA at 0, 10, 20 and 50 nM. p Quantitative analysis of GSDMD oligomer/GSDMD monomer. q Non-reducing immunoblotting of GSDMD oligomer. Murine BM neutrophils were pretreated with IMT1 or CsA plus VBTI4 and then treated with IFN-γ plus LS. r Quantitative analysis of GSDMD oligomer/GAPDH. n = 5 (r) or 6 mice (b, d, f, h, j, l). Plots were pooled from five independent experiments (n, p). The immunoblotting samples shown are from the same experiment. Three blots were processed in parallel (b, d, f, h, j, i, r). Data are presented as mean ± SD. Significance was examined with one-way ANOVA (b, f, h, j, l, n, p, r) or unpaired two-sided Student’s t test (d).