Inflammasome activation results in the generation of ox-DNA. (A) Immortalized murine SGM model cell lines (Srsf2^P95H and Tet2^−/−) display increased activity of caspase-1 assessed by Caspase-1 Glo assay in mutant compared with WT cells (mean ± standard deviation [SD] of 3 independent experiments). (B) Representative image of increased intracellular ox-DNA assessed by flow cytometry (mean fluorescent intensity [MFI] ± SD of 3 independent experiments) in immortalized murine SGM model cell lines (Srsf2^P95H and Tet2^−/−) compared with WT controls. (C) Confocal IF of immortalized murine SGM models stained for ox-DNA (4′,6-diamidino-2-phenylindole [DAPI]; ox-DNA/RNA; original magnification ×2520). Micrographs are representative figures. MFI of cells was quantified (minimum 200 cells per sample, 3 samples per group; MFI ± SD). (D) Cell-free ox-DNA was assessed by ELISA in media supernatants of immortalized SGM cell lines (mean ± SD of 3 independent experiments). (E) Confocal IF of cytosolic ox-mtDNA in MDS BM-MNC samples compared with normal BM-MNC (DAPI, ox-mtDNA fluorescein isothiocyanate [FITC]; original magnification ×2520). Representative micrographs. MFI of cells was quantified (minimum 200 cells per sample, 3 samples per group; MFI ± SD). FL1-H, fluorescence parameter 1 high.