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. 2001 Feb 1;29(3):668–676. doi: 10.1093/nar/29.3.668

Figure 6.

Figure 6

Partial rescue of mtDNA amount and integrity of AS1 cells by overexpression of mitochondrial DNA ligase III. (A) Western blot analysis of mitochondrial extracts prepared from control cells, AS1 cells and AS1 cells overexpressing mitochondrial DNA ligase III. Aliquots of 10 mg mitochondrial extracts were separated by 10% SDS–PAGE and transferred to a nylon membrane. Blots were probed with anti-ligase III antibody at a 1:500 dilution. (B) The amount of mtDNA in control, AS1 cells and AS1 cells overexpressing mitochondrial DNA ligase III was measured. Genomic DNA digested with PvuII was separated on a 0.5% agarose gel under native conditions, transferred to a nylon membrane and probed with a portion of mtDNA. The mtDNA was quantified using IP Labgel. Three independent experiments were carried out and the average value obtained plotted as a graph. Standard errors of the mean are represented as error bars. (C) PvuII-digested genomic DNA from control cells, AS1 cells and AS1 cells overexpressing mitochondrial DNA ligase III was denatured in the presence of 0.2 N NaOH prior to electrophoresis on a 0.5% agarose gel. The blot was treated similarly to as described above.