Activation of the cGAS - STING pathway in VPS13C ^KO cells is dependent on increased cytosolic mtDNA. (A) qPCR of three ISG transcripts shows that depletion of mtDNA with EtBr reduces ISG levels in VPS13C^KO cells to or near WT levels. n = 3 biological replicates. (B) p-TBK1 and p-STING are reduced in EtBr-treated VPS13C^KO cells. (C) Levels of three mtDNA amplicons (ND1, D-Loop, and CYB) are elevated in the cytosolic fraction of VPS13C^KO cells detected by qPCR and normalized to nuclear gene hB2M. n = 3 biological replicates. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001. #, P < 0.05; ##, P < 0.01; ###, P < 0.001; #### < 0.0001 in EtBr compared with untreated cells. Data were compared using two-sided t tests. Error bars represent ±SD. Source data associated with this figure can be found at https://doi.org/10.5281/zenodo.6416363. Source data are available for this figure: SourceData F4.