Extended Data Fig. 10: Loss of Snx9 rescues mtDNA release and TBK1–IRF3 activation triggered by Fh1 loss.
From: Fumarate induces vesicular release of mtDNA to drive innate immunity
a, Representative confocal images of mitochondrial morphology (TOM20) and DNA foci (DNA) in cFh1−/−CL19 cells transfected with scramble (scr) or Snx9 siRNA. White arrows indicate cytosolic DNA foci. Scale bars: 10 μm. b, Number of cytosolic DNA foci per cell in cFh1 cells. n = 3 independent experiments. c, Percentage of cells with cytosolic DNA foci in cFh1 cells. n = 3 independent experiments. d, Immunoblots of specified proteins of cFh1 cells. e,f, Quantification of mitochondrial number (e), and area (f), in cFh1 cells. n = 3 independent experiments. g,h, Average number of cytosolic DNA foci per cell (g), and number of cytosolic DNA foci per cell (h), in iFh1fl/flCL29 cells transfected with scr or Snx9 siRNA and treated with either vehicle (EtOH) or 4-OHT (iFh−/−CL29) for the indicated period of time. n = 3 independent experiments. i, Immunoblots of specified proteins in iFh1CL29 cells. j-l, Quantification of mtDNA copy number by ddPCR using either a mtCo3 (j), ND1 (k) or D-loop (l) probe, from isolated cytosolic fractions of iFh1fl/flCL29 cells. n = 3 independent experiments. Data are mean ± s.e.m. b,c,e-h,j-l, one-way ANOVA with Tukey’s multiple comparison test.
