Extended Data Fig. 4: Additional data supporting the role of PARP9 in mtDNA-stress-mediated enhancement of nDNA damage responses.
From: Mitochondrial DNA stress signalling protects the nuclear genome
a, Heat map of PARP family gene expression from previously published microarray data4 in WT and Tfam+/- littermate MEFs (two of each). b, RT-qPCR analysis of Parp9 expression in WT (Tfam+/+ Stat1+/+), Tfam+/-, Stat1-/- and Tfam+/- Stat1-/- littermate MEFs. c, RT-qPCR analysis of Parp9 expression at the indicated times after transfection with two Parp9 siRNAs (#1 and #2). b and c, the data shown are from one of three biological replicates with the error bars indicating the mean ± s.d. of three technical replicates. The other two biological replicates are provided as Supplementary Figure 6. d, Tfam+/- MEFs were transfected with control (Ctrl) or one of two Parp9 siRNAs (#1 and #2) for 48 h and then assessed for cell viability using the alamarBlue assay after treatment with 1.0 or 1.5 µM doxorubicin (Dox) for 24 h. Error bars indicate mean ± s.d. of n=4 biological replicates. e–h, Analysis of DNA repair rate (that is the rate of γH2A.X and p-53BP1 foci resolution during recovery after (e, f) 2 Gy IR or (g, h) 12 h of 1µM doxorubicin-mediated damage. Nuclei are labelled with DAPI (blue), while γH2A.X (magenta), and p-53BP1 (green) were detected by immunofluorescence. (e–h) Plotted to the right of the images is the average number of foci per nucleus at the indicated times. Scale bars represent 15 µm. Error bars indicate means ± s.d. of n=3 biological replicates, with 50 nuclei quantified in each. All data were analysed with two-tailed unpaired student’s t tests. Asterisks indicate significance as follows: * P < 0.05, ** P < 0.01, *** P < 0.001.
