Extended Data Fig. 3: Chronic loss of Fh1 leads to abnormal mitochondrial morphology and mtDNA release in the cytosol.
From: Fumarate induces vesicular release of mtDNA to drive innate immunity
a, Representative N-SIM super-resolution images of mitochondrial morphology (TOM20) in epithelial kidney cell line with chronic Fh1 deletion (cFh1−/−CL1) from cFh1fl/fl, compared to cFh1fl/fl cells. Scale bar: 5 μm. b, Quantification of mitochondrial diameter in cFh1 cells from (a). n = 3 independent experiments. c, Quantification of mitochondrial morphology in cFh1 cells. n = 3 independent experiments. d, Quantification of mitochondrial number (left) and area (right) in cFh1 cells. n = 3 independent experiments. e, Representative confocal images of mitochondrial morphology (TOM20) and DNA foci (DNA) in cFh1 cells. White arrows indicate cytosolic DNA foci. Scale bar: 10 μm. f,g, Percentage of cFh1 cells showing cytosolic DNA (f), and number of cytosolic DNA foci per cell (g), from e. n = 6 independent experiments. h-j, Quantification of mtDNA copy number by ddPCR using either a mtCo3 (h), ND1 (i) or D-loop (j) probe, from isolated cytosolic fractions of cFh1 cells. n = 3 independent experiments. k, Immunoblots of specified proteins in cFh1 cells. l, Expression of inflammation-related Ifi202b and ISGs in cFh1 cells measured by qRT-PCR. n = 3 independent experiments. Data are mean ± s.e.m. b,d,f-j, one-way ANOVA with Tukey’s multiple comparison test, c,l, two-way ANOVA with Tukey’s multiple comparison test.
