Extended Data Fig. 6: Ongoing mtDNA replication is required for ISG expression.
From: Ribonucleotide incorporation into mitochondrial DNA drives inflammation
a, RT-qPCR analysis of wild-type MEFs depleted of replication enzymes with the indicated siRNAs for 72 h. mtDNA levels were monitored with probes for D loop, cytochrome b (CytB) and non-NUMT (non-nuclear mitochondrial DNA). n = 3 biologically independent experiments. b, ISG expression in Mgme1+/+ and Mgme1−/− immortalized MEFs treated with the indicated siRNA or esiRNA for 72 h. n = 3 independent cultures. c, A representative immunoblot analysis with the indicated antibodies in Mgme1+/+ and Mgme1−/− immortalized MEFs treated with the indicated siRNA for 72 h. d, Ribonucleotide levels in cell lysates of Mgme1+/+ and Mgme1−/− immortalized MEFs, determined by LC-MS (n = 6 independent cultures). Samples were normalized to protein levels. e, Ratio of ribonucleotides triphosphates (rNTP) and deoxyribonucleotides triphosphates (dNTP) from cell extracts of immortalized Mgme1+/+ and Mgme1−/− cells treated with control and siRNAs for Samhd1, determined by LC-MS (n = 6 independent cultures). P values were calculated using two-way ANOVA with Tukey’s (a, e) or Šidák’s multiple comparison test (d) or one-way ANOVA with Tukey’s multiple comparison test (b). data are presented as mean ± SD.
