Cytosolic release of mtDNA in iNgly1 ^−/− mouse cerebellum. (A) Immunoprecipitation of endogenous cGAS in the Ngly1^fl/fl, iNgly1^−/− cerebellar lysate. n = 3 mice per genotype. (B) qPCR analysis of cGAS-bound mtDNA in the Ngly1^fl/fl, iNgly1^−/− cerebellar lysate. n = 3 mice per genotype. Data are the mean ± SEM. Unpaired Student’s t test. **P < 0.01. (C) Quantification of cholesterol in Ngly1^fl/fl, iNgly1^−/− cerebella. n = 3 mice per genotype. Data are the mean ± SEM. Unpaired Student’s t test. *P < 0.05. (D) Quantification of cholesterol in SK-N-SH cells treated with simvastatin at indicated concentrations for 24 h. Data are the mean ± SEM of three biological replicates. Unpaired Student’s t test. ***P < 0.001. (E) qRT-PCR analysis of IFNB1 gene expression in simvastatin-treated SK-N-SH cells after with or without diABZI (2 µM) treatment for 2 h. Data are the mean ± SEM of three biological replicates. Unpaired Student’s t test. *P < 0.05; ns, not significant. (F) Quantification of cholesterol in SK-N-SH cells treated with cholesterol-depleting agent MβCD at indicated concentrations for 48 h. Data are the mean ± SEM of three biological replicates. Unpaired Student’s t test. ***P < 0.001. (G) qRT-PCR analysis of IFNB1 gene expression in MβCD-treated SK-N-SH cells after with or without diABZI (2 µM) treatment for 2 h. Data are the mean ± SEM of three biological replicates. Unpaired Student’s t test. *P < 0.05; ns, not significant. Source data are available for this figure: SourceData FS5.