Fig. 2

a (a-1) DNase II knockdown (6 days after transfection) in SH-SY5Y cells. siRNA-mediated knockdown was confirmed at the mRNA and protein (western blotting) levels. a-2 Immunostaining for dsDNA, histone H2B, and Hsp60 in SH-SY5Y cells transfected with the DNase II siRNA shows cytosolic dsDNA deposits (white arrow). The bar graph shows the ratio of ectopic dsDNA+ cells among SH-SY5Y cells transfected with DNase II siRNAs. a-3 The death of DNase II-depleted cells was assayed by immunofluorescence staining for caspase-3. b (b-1) In situ hybridization of mitochondrial DNA and coimmunostaining for histone H2B and Hsp60 in SH-SY5Y cells transfected with DNase II siRNA. A white arrow indicates cytosolic dsDNA of mitochondrial origin. b-2 Transmission electron micrographs of SH-SY5Y cells transfected with the DNase II siRNA. c qPCR analysis of DNase II, IL-1α, IL-1β, IL-6, IL-8, and IFN-β mRNAs in DNase II-depleted SH-SY5Y cells. N.S.: statistically not significant. d Effects of DNase II overexpression on the death of SH-SY5Y cells with GBA, ATP13A2, and PINK1 knockdown. d-1 The increase in cytosolic dsDNA deposits in GBA-, ATP13A2-, and PINK1-depleted SH-SY5Y cells were suppressed by DNase II overexpression. si-Triple: Knockdown of GBA, ATP13A2, and PINK1 expression with siRNAs. d-2 Cell death and rescue effects were assayed using western blotting for cleaved Caspase-3 and cleaved Gasdermin D and propidium iodide (PI) staining. si-Triple: Knockdown of GBA, ATP13A2, and PINK1 expression with siRNAs. e Effects of DNase II overexpression on the type I IFN responses of SH-SY5Y cells with GBA, ATP13A2, and PINK1 knockdown. si-Triple: Knockdown of GBA, ATP13A2, and PINK1 expression with siRNAs. The statistical details are described in Supplementary Table 4. Source data of Fig. 2 are provided as a Source Data file.