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. 2023 Nov 30;14:7889. doi: 10.1038/s41467-023-43635-y

Fig. 4. Depletion of mtDNA reduces ISG responses to iF17 infection.

Fig. 4

a, b NHDFs were treated with DMSO control or 25 μM ddC for 4 days. a Fixed cells were stained for DNA to confirm loss of mtDNA prior to infection. b RT-qPCR analysis of cytosolic fractions shows reduced mtDNA in the cytosol of iF17 infected cells treated with ddC. Samples are normalized to their respective mock to measure relative release as ddC reduces mtDNA levels overall. n = 3 per group, independent two-tailed t-tests. Data are presented as mean values ± SEM. c ddC treatment has a dose-dependent effect on ISG responses to iF17 infection but also reduces mitochondrial cytochrome b (mtCytb) levels in NHDFs. Molecular Weight markers are shown in kDa. d quantification of ISG expression in iF17-infected NHDFs treated with DMSO control or 25 μM ddC. n = 4 per group, independent two-tailed t tests. Data are presented as mean values ± SEM. e, f THP1 monocytes expressing an IRF reporter were treated with 10 μM ddC for 6 days prior to iF17 infection. e Western blot analysis of ISGs and viral proteins. Molecular Weight markers are shown in kDa. f Luciferase assays measuring IRF activity. n = 3 per group, 2-way ANOVA, Sidak’s multiple comparison test. Data are presented as mean values ± SEM. g Oxygen consumption rate (OCR) is reduced in 25 μM ddC-treated NHDFs. n = 9 per group, independent two-tailed t tests. Data are presented as mean values ± SEM. a–g Representative of 3 or more biological replicates. For panels (b, d, f, g); *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.0001. Source data are provided as a Source Data file.