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. 1997 Sep 15;186(6):921–929. doi: 10.1084/jem.186.6.921

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Inactivation of Ku70 by homologous recombination. (A) Diagrammatic representation of the Ku70 locus (top), the targeting construct (middle), and the targeted allele and hybridization probe (bottom). EcoRI (E) restriction sites used to detect the targeted gene are indicated (21). (B) Southern blot of EcoRI-digested tail DNA from control wild-type (WT), heterozygous (+/−), and homozygous (−/−) Ku70-targeted mice. The wild-type and mutant fragments are 13 and 5.7 kb, respectively. (C) Western blot analysis showing that Ku70 protein is not expressed in Ku70⁻ ^/− cells. Whole cell lysates prepared from mouse ear fibroblasts (50 μg) and mouse embryo fibroblasts (100 μg) were separated by 10% SDS-PAGE, transferred to a nitrocellulose membrane, and probed with polyclonal antibodies against full-length rodent Ku80 (top) and Ku70 (bottom), respectively. (D) Gel mobility shift assay (22) showing the lack of DNA-end–binding activity in Ku70⁻ ^/− cells. Ku-DNA–binding complex is indicated by arrow on the right.