Extended Data Fig. 7: Alum-induced Ox-mtDNA-dependent autoimmunity relies on Tfh cells.

a, Tfh frequencies after co-culture of murine Bcl6^ff and Bcl6^ΔCD4 naïve splenic CD4⁺ T cells with FACS-sorted B6 BM-Flt3L-induced pDC, −/+ Ox-mtDNA for 3 days (n = 3 biological replicates). b-j, Alum-induced autoimmunity in 6–8-week-old Bcl6^ff and Bcl6^ΔCD4 males treated as in Extended Data Fig. 3a. Intracellular BCL6 staining of splenic CD4⁺ T cells (b), serum anti-dsDNA IgG (c), FC analyses of splenic GC B frequencies (d), IF of GL7 (blue) and B220 (red) stained spleen sections (Scale bar, 50 μm, e), serum IL-21 and IL-1β (f), urinary protein ELISA (n = 4/group, g), sizes of glomeruli in kidney sections (n = 123/group, h), MFI of renal IgG deposits (n = 120/group) and C3 (n = 120/group) and renal area (in %) occupied by CD45⁺ cells (n = 29/Bcl6^ff PBS; n = 38/Bcl6^ff Alum; n = 33/Bcl6^ΔCD4 PBS; n = 32/Bcl6^ΔCD4 Alum) (i) and spleen mass to body weight ratio (j). k, Numbers and percentages of peritoneal pDC 24 h after PBS or alum injection into Bcl6^ff and Bcl6^ΔCD4 males (n = 5/Bcl6^ff PBS; n = 9/Bcl6^ff Alum; n = 5/Bcl6^ΔCD4 PBS; n = 8/Bcl6^ΔCD4 Alum). l, Active Casp1 (FLICA^hi) in pre-gated peritoneal pDC 24 h after alum injections into Bcl6^ff and Bcl6^ΔCD4 males (representative of 3 independent experiments). FMO, fluorescence minus one. Results in (a, c, d and f-k) are mean ± SD. n = 8/Bcl6^ff PBS; n = 7/Bcl6^ff Alum; n = 5/Bcl6^ΔCD4 PBS; n = 7/Bcl6^ΔCD4 Alum (b-d, f and j). Two-way ANOVA with Tukey multiple-comparison test. Micrographs (e) are representative of at least three independent experiments.

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