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. Author manuscript; available in PMC: 2021 Jul 28.
Published in final edited form as: Nat Immunol. 2021 Jan 28;22(3):312–321. doi: 10.1038/s41590-020-00859-0

Extended Data Fig. 2. Mitochondrial abnormalities in Irgm1−/− MEFs.

Extended Data Fig. 2

a,b, Interferon-β (Ifnb1) and interferon-stimulated gene (Ifit1) expression in mouse embryonic fibroblasts (MEFs) (n=3) (a) and bone marrow-derived macrophages (n=3) (b) treated with or without IFN-γ (20 ng/ml, 16h). c, Colocalization of dsDNA and HSP60 immunostaining expressed as Mander’s coefficient (n=4 for all conditions, except n=3 for Irgm1+/++IFN-γ). d, Mitochondrial gene Dloop1 quantified by qPCR in total DNA isolates of MEFs, normalized to nuclear gene Tert (n=7). e, Expression of mtDNA-encoded genes mt16S and mtND4 quantified by qPCR in MEFs (n=3). f, Mitochondrial fractions evaluated for purity by immunoblotting for mitochondrial protein TIM23 and cytosolic protein tubulin. g,h, qPCR of mtDloop1 (n=3) (g) and immunostaining for cytoplasmic dsDNA (h) confirming mitochondrial DNA depletion by EtBr (scale bar, 20 μm). i, Ifit1 expression in MEFs treated with mitochondria-specific antioxidant MitoTempo (50 μM) prior to IFN-γ (n=3). a, b and e-i are representative of at least two independent experiments. d is combination of two independent experiments. Data are mean +/− s.e.m. *P < 0.05, **P < 0.01, ***P < 0.001. Two-tailed unpaired t-test.