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. 2011 Aug 23;6(8):e23732. doi: 10.1371/journal.pone.0023732

Figure 6. Identification of t-circles in the ku70 null mutants.

Figure 6

(A) (Top) Genomic DNA samples were prepared from the parental wild type strain (BWP17) and its ku70-/- derivative (LCF2.1), digested with AluI and NlaIII, and then subjected to 2-D electrophoresis followed by Southern blotting to identify linear and circular telomeric DNA. (Bottom) The sample DNA samples were subjected to 2-D gel electrophoresis (without prior digestion) and analyzed using an rDNA probe. (B) Genomic DNA samples from the indicated strains were subjected to 2-D gel electrophoresis followed by Southern blotting to assess the levels of linear and circular telomeric DNA. All mutant strains were derived from the CAI4 parental strain, as follows: ku70-/- (LCD2A.1); ku70-/- lig4-/- (CEA2.5); ku70-/- rad52-/- (JLT2.1). (C) Genomic DNAs from the ku70-/- strain (LCD2A.1) were digested and analyzed by 2-D gel electrophoresis together with a nicked 4.4 kb circular plasmid (pGEM-URA3, isolated by gel electrophoresis). Radioactive probes for the plasmid and for telomere DNAs were both included in the hybridization mixture. A strip of DNA size standards corresponding to the first round of electrophoresis is shown at the bottom.