Extended Data Fig. 6. Post-stroke plaque inflammasome activation is mediated by cell-free DNA.

a. Representative EMSA gel microphotograph of different Calixarene concentrations (0–1000 µM) interfering with the AIM2-dsDNA complex resulting in increased free DNA. b. Quantification of AIM2-free DNA based on its relative fluorescence in the EMSA assay (K test; n = 3 per group; 3 independent experiments). c. ELISA analysis of IL-1β in CCA lysates from mice with tandem stenosis (TS), 24 h after stroke in control-, NLRP3 inhibitor- (MCC950) or AIM2 inhibitor- (4-sulfonic calixarene) treated mice, and in sham operated mice (K test; n = 5–6 per group). d. Total cell-free DNA (cfDNA), single-stranded DNA (ssDNA) and double-stranded DNA (dsDNA) in mouse serum 24 h after sham or stroke surgery (U test, n = 7–8 per group). e. Total DNA, ssDNA and dsDNA in mice serum after sham or 12 h, 24 h after myocardial infarction (MI) surgery (black: sham; blue circle: 12 h after MI; blue dot: 24 h after MI, multiple t test, n = 5–10 per group). f. Representative gel electrophoresis photographs of cfDNA isolated from sham and stroke-operated mouse plasma. g. Quantification of electrophoresis-based cfDNA fragment length analysis of sham or stroke-operated mice (K test; n = 4–5 per group; 0–400 bp fragments: Sham vs Stroke: P < 0.0001). h. Quantification of extra-vesicular and intra-vesicular DNA after sham or stroke surgery (U test; EV DNA: Sham vs Stroke P = 0.1746; vesicle-free DNA: Sham vs Stroke P = 0.0079; n = 5 per group).