Figure 2. Complex formation of CSA/CSB proteins with mtDNA as well as mtOgg1 and mtSSBP1 in oxidatively stressed mt. (A) GEMSA: Biotin-labeled mtDNA was incubated with mitochondrial extracts of oxidatively stressed normal or CSA-deficient human cells and with subsequent PAGE. Incubation with CSA antibody in WT mitochondrial extract showed a supershift (arrow). CoIP with biotin-labeled mtDNA. CoIP of CSA– (B) or CSB–mtDNA (C) complexes occurred only in WT mitochondrial extracts but vanished under DNase digestion. Complexes containing mtOgg1, mtSSBP1, COXIV, and ATP-Sβ were extracted by CoIP from mitochondrial extracts of CSA- and CSB-deficient and WT cells, exposed to 30 µM H2O2. The presence of CSA (D) and CSB (E) could be detected in complex with mtOgg1 and mtSSBP1 (arrows) by Western blot. Pictures are representatives of at least three independent experiments.
Figure 2.

Complex formation of CSA/CSB proteins with mtDNA as well as mtOgg1 and mtSSBP1 in oxidatively stressed mt. (A) GEMSA: Biotin-labeled mtDNA was incubated with mitochondrial extracts of oxidatively stressed normal or CSA-deficient human cells and with subsequent PAGE. Incubation with CSA antibody in WT mitochondrial extract showed a supershift (arrow). CoIP with biotin-labeled mtDNA. CoIP of CSA– (B) or CSB–mtDNA (C) complexes occurred only in WT mitochondrial extracts but vanished under DNase digestion. Complexes containing mtOgg1, mtSSBP1, COXIV, and ATP-Sβ were extracted by CoIP from mitochondrial extracts of CSA- and CSB-deficient and WT cells, exposed to 30 µM H2O2. The presence of CSA (D) and CSB (E) could be detected in complex with mtOgg1 and mtSSBP1 (arrows) by Western blot. Pictures are representatives of at least three independent experiments.

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