Extended Data Fig. 3: Alum-induced autoimmunity depends on NLRP3 and mtDNA oxidation.

a, Treatment and analysis scheme of WT & Nlrp3^−/− (b-d) and WT & mt-Ogg1^Tg (e-n) females. b-d, Relative amounts of circulating ccf-mtDNA (b), ratio of Fpg-treated (+) to nontreated (−) circulating ccf-mtDNA (c), and nDNA (Tert or B2m) to mtDNA (D-loop) ratio (d) (n = 8/WT PBS; n = 14/WT alum; n = 9/Nlrp3^−/− alum). e, IF of IgG (green), GL7 (blue) and B220 (red) stained spleen sections. Scale bar, 50 μm. f,g, Percentages of splenic GC B and Tfh cells (f) and splenic CXCR5⁻CXCR3⁺PD-1⁺CD4⁺ and IFNγ⁺IL10⁺CXCR5⁻CD4⁺ memory T cells (g) (n = 7/WT PBS, n = 10/WT alum, n = 7/mt-Ogg1^Tg alum). h, Percentages of blood CXCR5⁻CXCR3⁺PD-1⁺CD4⁺ T helper and IFNγ⁺IL10⁺CXCR5⁻CD4⁺ T memory (n = 6/WT PBS, n = 4/WT Alum, n = 4/mt-Ogg1^Tg Alum). i, Percentages of blood Tfh cells (n = 7/WT PBS, n = 10/WT Alum, n = 7/mt-Ogg1^Tg Alum) and CD138⁺CD19⁺CD3⁻CD4⁻ cells (n = 6/WT PBS, n = 4/WT Alum, n = 4/mt-Ogg1^Tg Alum). j, Glomeruli sizes in H&E-stained kidney sections (n = 165/WT PBS, n = 147/WT Alum, n = 158/mt-Ogg1^Tg Alum). k, MFI of IgG (n = 158/WT PBS, n = 166/WT Alum, n = 123/mt-Ogg1^Tg Alum), C3 deposits (n = 115/WT PBS, n = 119/WT Alum, n = 101/mt-Ogg1^Tg Alum) and area (in %) occupied by CD45⁺ cells (n = 74/WT PBS, n = 78/WT Alum, n = 72/mt-Ogg1^Tg Alum) in kidney sections. l, Urinary protein ELISA (n = 4/WT PBS, n = 7/WT Alum, n = 5/mt-Ogg1^Tg Alum). m,n, ELISA of serum anti-dsDNA and anti-nucleosome IgGs (m) and IL-1β (n) (n = 5/WT PBS or Alum, n = 6/mt-Ogg1^Tg PBS or Alum). Results in (b-d and f-n) are mean ± SD. Two-way ANOVA with Tukey multiple-comparison test. Micrographs (e) are representative of at least three independent experiments.

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